1 rhodopsin: effect of replacement of amino acids in the predicted folding core of rhodopsin growth...
Post on 03-Jan-2016
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Rhodopsin:Effect of replacement of amino acids in the predicted folding core of Rhodopsin Growth hormone receptor (GHR):Expression of Cytoplasmic domain of GHR4 in E.coli,
Epidermal Growth Factor (EGF):Secretory and extracelluar production of human EGF
Ciliary Neurotrophic Factor Alpha Receptor (CNTFR):Expression of Cytokine binding domain (CBD) of CNTFR
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Rhodopsin
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ResidueNameS22CR1F24CR2E25CR3A26CR4A27CR5N111CR6L112C/F116CdR7L112CR7F116CR8A166CR9A168CR10A169CR11
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Bovine Rhodopsin
GAATTCATGAACGGTACCGAAGGCCCAAACTTCTACGTTCCTTTCTCCAACAAGACGGGCGTGGTGCGCAGCCCGTTCGAGGCTCCGCAGTACTACCTGGCGGAGCCCTGGCAGTTCTCCATGCTGGCCGCCTACATGTTCCTGCTGATCATGCTTGGCTTCCCGATCAACTTCCTCACGCTGTACGTCACAGTCCAGCACAAGAAGCTTCGCACACCGCTCAACTACATCCTGCTCAACCTGGCCGTGGCAGATCTCTTCATGGTCTTCGGTGGCTTCACCACCACCCTCTACACCTCTCTCCATGGGTACTTCGTCTTTGGGCCGACGGGCTGCAACCTCGAGGGCTTCTTTGCCACCCTGGGCGGTGAAATTGCACTGTGGTCTCTGGTAGTACTGGCGATCGAGCGGTACGTGGTGGTGTGCAAGCCCATGAGCAACTTCCGCTTCGGTGAGAACCACGCCATCATGGGCGTCGCCTTCACCTGGGTCATGGCTCTGGCCTGTGCGGCCCCGCCGCTCGTCGGCTGGTCTAGATACATCCCGGAGGGCATGCAGTGCTCGTGCGGGATCGATTACTACACGCCGCACGAGGAGACCAACAATGAGTCGTTCGTCATCTACATGTTCGTGGTCCACTTCATCATCCCGCTGATTGTCATCTTCTTCTGCTATGGCCAGCTGGTGTTCACCGTCAAGGAGGCTGCAGCCCAGCAGCAGGAGAGCGCCACCACTCAGAAGGCCGAGAAGGAGGTCACGCGTATGGTTATCATCATGGTCATCGCTTTCCTAATCTGCTGGCTGCCCTATGCTGGTGTGGCGTTCTACATCTTCACCCATCAGGGCTCTGACTTTGGGCCCATCTTCATGACCATCCCGGCTTTCTTTGCCAAGACGTCTGCCGTCTACAACCCGGTCATCTACATCATGATGAACAAGCAGTTCCGGAACTGCATGGTCACCACTCTCTGCTGTGGCAAGAACCCGCTGGGTGACGACGAGGCGTCGACCACCGTCTCCAAGACAGAGACCAGCCAAGTGGCGCCTGCCTAAGCGGCCGC
MNGTEGPNFYVPFSNKTGVVRSPFEAPQYYLAEPWQFSMLAAYMFLLIMLGFPINFLTLYVTVQHKKLRTPLNYILLNLAVADLFMVFGGFTTTLYTSLHGYFVFGPTGCNLEGFFATLGGEIALWSLVVLAIERYVVVCKPMSNFRFGENHAIMGVAFTWVMALACAAPPLVGWSRYIPEGMQCSCGIDYYTPHEETNNESFVIYMFVVHFIIPLIVIFFCYGQLVFTVKEAAAQQQESATTQKAEKEVTRMVIIMVIAFLICWLPYAGVAFYIFTHQGSDFGPIFMTIPAFFAKTSAVYNPVIYIMMNKQFRNCMVTTLCCGKNPLGDDEASTTVSKTETSQVAPA
(Underlined nucleotide has been modified, opsin gene in PMT4 is different to the original sequence as shown above)
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Relative ratio A280/A500 of rhodopsin wide Type (WT) and 11 mutantsMutants F24C, A26C, L112C/F116C, F116C and A168C showed relative ratio between 500nm and 280nm were greater than 8Mutants S22C, E25C, A27C, N111C, A166C and A169C showed less than 5[Further experiments are still need to perform to further verify before the classified the mutants]
Chart18
2.30.33
4.170.61
19.112.36
3.840.27
9.360.96
4.150.69
2.540.21
20.195.24
14.143.75
3.10.25
7.740.84
2.530.12
Relative Ratio of A280/A500
Sheet1
WTR1R2R3R4R5R6R7R8R9R10R11
WTS22CF24CE25CA26CA27CN111CL112C/F116CF116CA166CA168CA169C
2.34.1719.113.849.364.152.5420.1914.143.17.742.53
0.330.612.360.270.960.690.215.243.750.250.840.12
Sheet1
00.33
00.61
02.36
00.27
00.96
00.69
00.21
05.24
03.75
00.25
00.84
00.12
Relative Ratio of A280/A500
Sheet2
Sheet3
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Elution profile of Rhodopsin mutant A169C first with NaPi buffer and then followed buffer with 150mM NaCl
Chart3
0.0042170.058021
0.0239170.087996
0.046720.107994
0.0266410.066425
0.0221810.05174
0.0183270.041144
0.0234360.056261
0.0542180.14752
0.0459720.110377
0.034230.080516
0.0294290.068123
0.0223780.05962
Absorbance 500 nm
Absorbance 280 nm
Fraction
Absorbance
Elution Profile of Rhodopsin Mutant A169C from 1D4 Sepharose Column
WT 1
6500.0018480.000083-0.000092-0.00211-0.001176-0.002988-0.0030930.0073640.0240430.0188040.0131450.0097770.0055250.005014
6490.0019130.000074-0.000078-0.002129-0.001258-0.003011-0.0032310.0076179310.0248720690.01945241380.01359827590.01011413790.00571551720.0051868966
6480.0019130.000037-0.000032-0.002065-0.001213-0.003011-0.003386totalper plate
6470.0018390.000051-0.000092-0.002102-0.001291-0.003053-0.0032270.08655724140.0173114483
6460.0019690.000133-0.00006-0.001951-0.001286-0.00298-0.003226
6450.0019830.000133-0.000041-0.002033-0.001204-0.003113-0.003209280-6500.037510.0489920.0361950.0277740.0232450.014280.014075
6440.002020.000120.000009-0.002061-0.001195-0.003063-0.003264500-6500.0073640.0240430.0188040.0131450.0097770.0055250.005014
6430.00190.000032-0.000097-0.002085-0.001287-0.003105-0.003292280/5005.09369907662.03768248551.92485641352.11289463672.37751866632.58461538462.807140008
6420.0019470.0000460.000028-0.002057-0.001236-0.003151-0.003324stdev 71.0986792164
6410.0019560.000041-0.000074-0.002108-0.001365-0.003192-0.003485stdev 60.343052336
6400.0019660.000083-0.000143-0.002044-0.001379-0.003316-0.003517error 70.415261711
6390.0020120.000055-0.00012-0.002072-0.001393-0.00328-0.003453error 60.1400505297
6380.0019340.000028-0.00017-0.002141-0.001407-0.00328-0.003545Ave of 72.7054866673
6370.0019620.000037-0.00023-0.002072-0.001481-0.003386-0.003642Ave of 62.3074512658
6360.001935-0.000028-0.000184-0.002132-0.001476-0.00346-0.00367
6350.001972-0.000129-0.000253-0.002188-0.001454-0.003461-0.003712
6340.002019-0.000005-0.00017-0.002183-0.001514-0.003571-0.003657
6330.001968-0.000129-0.000198-0.002138-0.001629-0.003649-0.003622
6320.001918-0.000124-0.000267-0.002212-0.001647-0.003673-0.00381
6310.002024-0.000055-0.000217-0.002166-0.001542-0.003673-0.00375
6300.002122-0.000023-0.000152-0.00213-0.001569-0.003751-0.003746
6290.0021170.000074-0.000074-0.00207-0.001634-0.003723-0.003696
6280.002034-0.000023-0.000157-0.002172-0.00163-0.003789-0.003839
6270.002057-0.000065-0.000157-0.00208-0.001607-0.003876-0.003972
6260.002160.000083-0.000157-0.002048-0.001634-0.003821-0.003858
6250.00216-0.000023-0.000115-0.002158-0.001644-0.003831-0.003927
6240.002132-0.000005-0.000175-0.002122-0.001658-0.00395-0.003918
6230.002184-0.000037-0.000157-0.002136-0.001612-0.003987-0.004001
6220.0022390.000074-0.000185-0.002118-0.00164-0.003882-0.003983
6210.0021790.000088-0.000208-0.002044-0.001663-0.003933-0.004097
6200.002272-0.000023-0.000203-0.002086-0.001668-0.003938-0.004025
6190.0022310.000014-0.000198-0.002031-0.001755-0.00397-0.004107
6180.0022730.000005-0.000226-0.002082-0.001741-0.004043-0.004085
6170.0022310.000023-0.000152-0.002091-0.001696-0.004062-0.004163
6160.0022680.000014-0.00012-0.002169-0.00176-0.004067-0.004223
6150.0023330-0.000166-0.002096-0.001719-0.004127-0.004085
6140.0022550.000111-0.000194-0.002022-0.001825-0.004159-0.004205
6130.0024540.000166-0.000005-0.001967-0.001737-0.004026-0.004017
6120.0024170.000134-0.000018-0.00199-0.001742-0.003999-0.004136
6110.002390.000092-0.000102-0.001995-0.001844-0.004211-0.004192
6100.0024040.000203-0.000051-0.001931-0.00171-0.004284-0.004129
6090.0023530.000125-0.000028-0.002001-0.001697-0.00417-0.004221
6080.0023950.0002080.000097-0.001936-0.001656-0.00423-0.004052
6070.002480.000217-0.000083-0.001955-0.001799-0.004323-0.004062
6060.0024390.0001940.000046-0.001974-0.001712-0.004173-0.00421
6050.0025040.0002590.000106-0.002034-0.00168-0.004183-0.00416
6040.0023050.0003290.000051-0.001851-0.001676-0.004179-0.004083
6030.0023790.0005180.000319-0.001703-0.001505-0.004065-0.004065
6020.0022790.0003660.000185-0.001806-0.001622-0.00421-0.004004
6010.0025310.0004170.000213-0.001774-0.001461-0.004005-0.00406
6000.0026810.0005650.000352-0.001687-0.001443-0.004089-0.00402
5990.0027520.0005750.000413-0.001674-0.001393-0.004041-0.003843
5980.0028650.0007330.000566-0.001508-0.001338-0.003922-0.003899
5970.0024980.0007010.000413-0.001565-0.001413-0.004008-0.00393
5960.0023040.000720.000465-0.001553-0.001405-0.004034-0.003906
5950.0024690.0008930.000632-0.001368-0.001211-0.003995-0.003875
5940.0025120.0009860.000623-0.001443-0.001174-0.003868-0.003772
5930.0025150.0010010.000694-0.001352-0.001208-0.004024-0.003632
5920.0025260.0010250.000676-0.001265-0.001223-0.003949-0.003741
5910.0024810.0010260.000863-0.001372-0.001159-0.003952-0.003739
5900.0022920.0010090.000775-0.001304-0.001244-0.004086-0.003845
5890.0022230.0011220.000822-0.001282-0.001226-0.004061-0.00388
5880.0020650.0012960.00087-0.001278-0.001241-0.003994-0.003777
5870.002010.001470.001015-0.001125-0.001032-0.003969-0.003788
5860.0019790.0014860.001082-0.001159-0.001117-0.003856-0.003722
5850.0018060.0015660.001242-0.001047-0.001089-0.004026-0.003882
5840.0015910.0016520.001196-0.001156-0.001151-0.003979-0.003993
5830.0016060.0016910.001432-0.001063-0.00104-0.004018-0.003865
5820.0018240.0019890.001433-0.000928-0.001031-0.003922-0.003927
5810.0019290.0018810.001533-0.001031-0.000881-0.004027-0.003962
5800.0017930.0021270.001717-0.000849-0.000872-0.003935-0.003856
5790.0019490.002270.001671-0.000831-0.000859-0.003942-0.004021
5780.0019120.0023840.001865-0.000812-0.000709-0.003901-0.003748
5770.0016480.0025740.002002-0.0007-0.000765-0.00387-0.003739
5760.0019360.0027210.00222-0.000573-0.000592-0.003823-0.003777
5750.0021020.0029250.002244-0.000503-0.000644-0.003853-0.00382
5740.0024610.0031380.002608-0.000348-0.000371-0.003718-0.003568
5730.0027130.00330.002741-0.000343-0.000442-0.00376-0.003648
5720.0031010.0035650.002722-0.000089-0.000348-0.003685-0.003676
5710.0032810.0036410.003091-0.000038-0.000244-0.003648-0.003476
5700.003210.0038740.0030630.000052-0.000212-0.003523-0.003541
5690.0033140.0040680.0033140.000273-0.00008-0.003518-0.003532
5680.003390.0042350.0034140.000263-0.000099-0.003537-0.003415
5670.0034140.0045770.0036270.0003390.000075-0.003485-0.003336
5660.0033670.0046680.0037790.0003860.000094-0.003528-0.003458
5650.0035270.0050140.0039310.0005980.000263-0.003438-0.003438
5640.003670.0051340.0041020.0007010.000311-0.003346-0.003374
5630.0036650.0054470.0045190.000880.000598-0.003251-0.003237
5620.0037070.0058370.0046710.0010120.000645-0.002994-0.003153
5610.0037260.0060230.0048610.0010830.000758-0.003144-0.003139
5600.0037070.0062810.0050610.0010730.000819-0.003135-0.002948
5590.0039430.0067790.0053730.0015780.001054-0.003035-0.002998
5580.0040830.0070710.0057390.0016970.001145-0.002682-0.002813
5570.0041460.0075470.0061220.0020470.001447-0.002641-0.002632
5560.0042420.0077960.006280.0021460.001504-0.002637-0.00266
5550.0042130.0080410.0065620.0022930.001674-0.00251-0.00252
5540.004470.0084860.0068150.0025010.001759-0.00237-0.002468
5530.0044740.0088160.0070770.0026760.002108-0.002178-0.00222
5520.0046160.0093340.0074350.0029030.002113-0.002098-0.002163
5510.0046870.0096080.0078460.0030830.002335-0.001999-0.002201
5500.004830.0100310.007980.0032440.002392-0.00192-0.002084
5490.004910.0104010.0082910.0034520.002675-0.001779-0.001952
5480.0050620.0106660.008660.003680.002732-0.001784-0.001877
5470.0050570.0110520.00890.0038220.002808-0.001624-0.001863
5460.0051420.0116090.0092770.0041340.003176-0.001408-0.001629
5450.0051750.0119130.0095560.0043480.003176-0.001277-0.001451
5440.0052980.0123420.0098340.0045750.003484-0.001239-0.001371
5430.0053220.0127670.0102370.0048080.003687-0.001033-0.001361
5420.0054780.0131730.0104340.0049930.003777-0.000939-0.001155
5410.0055070.0136140.0108820.0053070.004067-0.00078-0.001094
5400.0056020.0139590.0111380.0054930.00429-0.000653-0.00085
5390.0058220.0144040.0114970.0057030.004367-0.00054-0.000817
5380.0056810.0147130.0116730.0058520.00452-0.000437-0.000733
5370.005890.0152390.0121420.0061530.004734-0.000273-0.000611
5360.0061760.0156660.0123730.0063760.004985-0.000094-0.00039
5350.0061580.0160390.0126990.0065970.0051290.000005-0.000296
5340.0062330.0164420.0131090.0070160.0054040.000395-0.000183
5330.0062730.0169050.0134030.0069990.0054670.000306-0.000169
5320.0063540.01720.0136810.0072380.0056820.000386-0.000061
5310.0063690.0175820.0138950.0074350.0057580.0005040.000151
5300.0065420.0179770.0142520.0076080.0059360.0006260.000151
5290.0066040.0182530.0145250.0077710.0060980.0005930.000249
5280.0068080.0188730.0147940.0080670.0063160.0009090.000329
5270.0069440.0190690.0149440.0081980.0063660.0009850.000311
5260.0068910.0193940.0151640.0083610.0064660.0009850.000504
5250.007050.0196310.0153840.0087220.0066770.0011310.000541
5240.0071550.0201190.0156820.008650.0067630.001220.000584
5230.0072810.0203750.0159140.0087050.0069120.001240.00073
5220.0073580.0206370.0161740.0089830.0071040.0012870.000674
5210.0073490.0208720.0163470.0091820.0071910.0014240.000782
5200.0075220.0212190.0165090.0092980.0072820.0015140.000933
5190.0075550.0214760.0167820.0093940.007320.0016040.00098
5180.007860.0217980.0171170.0096760.0076110.001840.001202
5170.0081350.0220240.0172120.0098550.0076790.0018260.001339
5160.0083930.0222790.0174110.0099940.0079760.0020330.001315
5150.0083560.0224630.0174550.0100060.0078530.0019770.001438
5140.008510.0226210.0176350.0102080.0078720.0020290.001415
5130.0084680.0227640.0176580.0101080.0080410.0019020.001254
5120.0086070.0230160.018030.0103390.008060.0021050.001377
5110.0085640.0230610.0179080.0103690.008070.0020530.001486
5100.0085590.0232110.0181050.0103550.0082040.0020770.001552
5090.0086190.0233450.0180050.0105010.0080910.0021390.001571
5080.0087710.0235050.0182040.0105570.0082910.0022470.001547
5070.0087570.0236650.0183320.0107070.008430.0023180.001651
5060.0087830.0236170.0184060.0106720.0083370.002210.001406
5050.0088790.0238050.0184550.0107580.0084420.0023050.001666
5040.008830.0237050.0184080.0106080.0082150.0021820.001548
5030.0089870.0240080.0184430.0108230.0085350.0024280.001633
5020.0089630.0240810.0186020.0109060.008430.0022910.001733
5010.0089990.0240370.018430.0108170.008350.0022960.001776
5000.0092120.0241260.0187120.0110350.0086010.0025370.001921
4990.0092010.0240620.0186660.011020.0083360.0023670.001799
4980.0091630.024150.0186850.0108890.0085520.0024670.001785
4970.009170.0239070.0184880.0109640.0084480.0023630.001724
4960.0093270.0240090.0185740.0109580.0085050.0023630.001676
4950.0092280.0239630.018390.010790.008520.0024960.00162
4940.0092570.0238840.0184450.010830.0084730.0023440.001752
4930.0093150.0238410.0182640.010830.0084160.0022970.001682
4920.0094020.0236710.018210.0108250.0083770.0023070.001606
4910.0093320.0236210.0181590.0106730.0082730.0021890.001653
4900.0092040.0235540.0180170.0107240.0082850.0021470.001573
4890.0093350.0233520.0179990.0106380.0082760.0021140.001436
4880.0092780.0232940.0178290.0104990.008070.0021520.001446
4870.0095630.0232560.0176780.0105530.007970.0022280.001413
4860.0096740.0230560.0178440.0104750.0081520.0021750.001484
4850.0094870.0229350.0176360.0104910.0081050.0020380.001413
4840.009490.0226830.0174050.0102870.0078530.0019110.001271
4830.0095010.0225280.0172610.0102010.0078640.0018590.001281
4820.0095490.0223870.0171560.0101760.0077960.001920.00121
4810.0095590.0221750.0169660.0101050.0076720.0018120.00121
4800.0095160.0220520.0168190.0100420.0077010.0017310.001106
4790.0094510.021640.0165140.0096820.0074140.0015750.001026
4780.0094030.0214820.0163430.0095810.0073660.0015330.001064
4770.0093940.0212460.0161050.0094810.0071130.0014090.001002
4760.0095140.021160.0161180.0095140.0071260.0013760.000936
4750.009510.0208150.0158890.0093210.007060.0013860.000865
4740.0095830.0206870.0157230.0091970.0070410.0013430.000714
4730.0093640.0202840.0153130.0089540.0067080.0010410.000553
4720.0095710.0201140.0152480.0089530.0066980.0010450.000619
4710.0094430.0197540.0149490.0087780.0064690.0009130.000487
4700.0095580.0196420.0147860.0086370.006310.000880.000463
4690.0094440.0192960.0146130.0083930.0062970.0006760.000359
4680.0094060.018940.0142760.0083690.0061250.0005530.000307
4670.0094090.0187290.014220.0082130.0060420.0005960.000331
4660.0095240.0185350.0138720.0080480.0059650.0004820.000175
4650.0092830.0181580.0136880.0079290.0057830.0003070.000095
4640.0095130.0179920.0135150.0077830.0057250.000180.000024
4630.0093210.0175660.0132290.0076210.005529-0.000005-0.000184
4620.0092960.017270.0129650.0075140.0053950.000014-0.000222
4610.0094180.0171040.0127810.0073470.005242-0.000123-0.000444
4600.0093890.016780.0125530.007040.005171-0.000264-0.000416
4590.0093980.0165240.0124210.0070730.004955-0.000255-0.000486
4580.0095050.0161870.0120870.0069680.004889-0.000368-0.000529
4570.0094430.0160550.0119080.0067660.004784-0.000482-0.000534
4560.0094680.0157870.0117920.0066280.004699-0.000397-0.000713
4550.0094390.0154550.0115940.0064460.004622-0.000652-0.000666
4540.0094650.0152510.0113630.006480.004551-0.000699-0.000586
4530.0095510.014920.0111530.0062720.004319-0.000742-0.000775
4520.0094780.0147140.0109250.0061810.004295-0.000709-0.000808
4510.009430.0143550.0106020.0059830.004159-0.000921-0.000888
4500.0093970.0140990.0103890.0057940.003951-0.001045-0.001068
4490.0095290.0138960.010410.0057320.004057-0.001017-0.000946
4480.0095890.0136220.0100390.0057270.003692-0.001225-0.001168
4470.0092740.0131770.0097740.005320.003456-0.001392-0.001354
4460.0095020.0130420.0095840.0052510.003425-0.001473-0.00136
4450.0093930.0128170.0093970.0050570.003332-0.001559-0.001559
4440.0094820.0125620.0091660.0049840.003276-0.001674-0.001598
4430.0095030.0122130.0090120.00480.003125-0.00176-0.001561
4420.0094120.0121190.0088480.0047840.002898-0.0017-0.001799
4410.0095180.0118250.0086040.004670.00287-0.001852-0.001848
4400.0094520.0115360.0084750.0043990.00268-0.002204-0.001901
4390.0094570.0112970.0081630.004190.002687-0.00222-0.002168
4380.0095190.0109450.0079240.0041340.002515-0.002268-0.002154
4370.0093770.0108470.0076990.0038950.002224-0.002416-0.002355
4360.0095370.0106310.0076780.0039980.002138-0.002621-0.002346
4350.0095910.0103780.0073910.0036820.002029-0.002689-0.002466
4340.0094780.0102640.0073350.0035770.001943-0.002813-0.002448
4330.0097130.0101620.0072140.0036260.001982-0.002723-0.002372
4320.0095910.0099520.0070480.0035020.001781-0.002701-0.002487
4310.0096570.0095990.0068030.0033880.001782-0.002882-0.002631
4300.009670.0095480.0067170.0033020.001571-0.002869-0.002612
4290.0095070.0091750.0064670.0030960.001577-0.003103-0.002903
4280.0096020.0090250.0063760.0030250.001452-0.00327-0.002994
4270.0097090.008830.0061140.0029520.001471-0.003198-0.002828
4260.0097240.0087910.0061330.0029470.001346-0.003336-0.002904
4250.0096110.0085110.0059760.0027950.001131-0.003508-0.003152
4240.0097950.0084160.0057160.0025260.001059-0.003576-0.003196
4230.0097470.0082460.0056720.0024630.001021-0.0035-0.003153
4220.0098870.0081510.0055910.0026180.000964-0.003435-0.003145
4210.0099070.0080490.0054950.0025030.00106-0.00353-0.003131
4200.0099020.0079030.0053740.0024260.000935-0.003597-0.003207
4190.0100790.0079120.0053980.0024350.000973-0.003582-0.003216
4180.0099310.0076380.0051480.002330.00083-0.003688-0.00337
4170.0101420.0076280.005090.0022720.000897-0.003783-0.003161
4160.0101570.0075310.0050660.0022150.000873-0.00375-0.003232
4150.0101970.0074480.0049550.0023160.000801-0.003788-0.003308
4140.010260.0073050.0048740.0020860.000715-0.003861-0.003295
4130.0102670.0071940.0047590.0020280.000475-0.003971-0.003367
4120.0102390.0070010.0046240.0019470.000485-0.0041-0.003568
4110.0103830.0069280.0046240.0019320.000595-0.004006-0.003406
4100.0103520.0068820.0044850.0019280.000451-0.004069-0.003569
4090.0105530.0069160.0045580.0019280.000456-0.003964-0.003436
4080.0104860.0068190.0044660.0019620.000538-0.00405-0.003384
4070.0106580.0066960.0043760.0017510.000432-0.004066-0.003567
4060.010740.006880.0044870.0019440.000605-0.003952-0.003423
4050.0107240.0067750.0044840.0019680.000428-0.004196-0.003525
4040.0107420.0066010.004170.0017080.000341-0.004202-0.003649
4030.0108030.0065250.0042820.001680.000399-0.004279-0.003603
4020.0108730.0065790.0041910.0017910.00038-0.004265-0.003655
4010.0108810.0065270.0042210.0017140.000317-0.004314-0.003737
4000.010880.0064420.0040670.0015650.000289-0.004367-0.003729
3990.0110270.0064680.0040880.0015510.000226-0.004354-0.003797
3980.0110870.0065660.0042530.0017160.000241-0.004302-0.003731
3970.0113050.0065520.0041760.0017590.000395-0.00435-0.003779
3960.0113730.0065840.0041190.00160.000313-0.004238-0.003603
3950.0113830.0065130.0040620.0016450.000347-0.004239-0.003777
3940.0112910.0065880.0040870.0017270.00027-0.004292-0.003672
3930.011490.006520.0040390.0016640.00026-0.004302-0.003706
3920.0116220.0066150.0041180.0016790.000284-0.00437-0.003702
3910.0114770.006490.0041930.0016560.000308-0.004391-0.003723
3900.0117810.0066770.0041450.0018640.000434-0.004349-0.003762
3890.0117760.006630.0041170.001710.000439-0.004436-0.003792
3880.011810.0064620.0040750.0016720.000328-0.004481-0.003812
3870.0119480.0066470.0041820.0018410.000323-0.004333-0.003635
3860.0120850.0066980.0041630.0017930.000376-0.004267-0.003708
3850.0120380.0067190.0041150.001740.000391-0.004344-0.003733
3840.0123630.00680.0042740.0017770.000498-0.004294-0.003638
3830.0123660.0067830.0040010.0019030.000312-0.004298-0.003631
3820.0123180.0067860.004120.0018130.000327-0.004376-0.003656
3810.0125250.006820.00430.0019370.000502-0.004282-0.003375
3800.0126780.006910.0042440.0018530.000487-0.004063-0.003566
3790.0126360.0069170.0041420.0018440.00039-0.004435-0.003606
3780.0128490.0071270.004370.0020010.000729-0.004062-0.003253
3770.0128260.0072060.0043920.0020590.000622-0.00417-0.003352
3760.012810.0071270.0043230.0019020.00051-0.004136-0.003482
3750.0130660.0072380.0044640.0021310.000539-0.004091-0.003452
3740.0131950.0070620.0044860.0020960.000655-0.004008-0.003493
3730.0132060.0073410.004480.0022120.00067-0.004141-0.003478
3720.0134770.0073520.0044210.0021050.00068-0.004252-0.003444
3710.0133450.0074150.0044650.0020610.000582-0.004141-0.003526
3700.0134750.0076040.0046450.002250.000708-0.004072-0.003467
3690.0135460.0074520.0045910.0021920.000815-0.004106-0.003342
3680.0136760.007580.0046990.0021720.000767-0.004034-0.003385
3670.0137740.0076830.0046050.0023140.000786-0.004082-0.003438
3660.0140310.0077790.0047460.0024150.000897-0.003999-0.003307
3650.013870.0076630.0047470.002260.000723-0.00401-0.003313
3640.014140.0079270.0048090.0023810.001053-0.004003-0.003302
3630.0142570.0079530.004790.0024590.000975-0.003985-0.003249
3620.0145230.0080640.0050630.0025560.001053-0.003753-0.003162
3610.0144660.0080850.0049470.0025810.001101-0.003859-0.003086
3600.0146190.0080170.0049820.0025080.001092-0.003918-0.00324
3590.0145940.008190.0050260.0025910.001048-0.003884-0.003048
3580.0148120.0082530.0051380.0026930.001204-0.003816-0.002965
3570.0149270.0083670.0051180.002830.00134-0.003653-0.002965
3560.0151720.0084580.0052130.0027520.001204-0.003726-0.002947
3550.0150660.0084310.0050920.0028160.001209-0.003693-0.003001
3540.0152730.008550.0052490.0029240.001282-0.003646-0.002803
3530.0154350.0087190.0055290.0029340.001477-0.003458-0.002741
3520.0153170.0085650.0051820.0028010.00122-0.003828-0.003067
3510.0155770.0085030.0053060.0029330.001328-0.003704-0.002904
3500.0156910.00870.0054070.0029590.001372-0.003657-0.002968
3490.0158240.0089230.0055520.002990.001568-0.003457-0.002641
3480.0157630.0085120.0052880.0028860.00115-0.003924-0.003016
3470.0159710.0088330.0053390.0030140.00137-0.003564-0.002781
3460.016060.0087390.0053850.0029640.001474-0.003703-0.002764
3450.0160840.0087060.0054490.003040.001368-0.003653-0.002849
3440.0163340.0087150.0052810.002910.00134-0.003705-0.002973
3430.0165810.0089360.0055490.0030530.001438-0.003478-0.002814
3420.0166980.008780.0055460.0030990.001596-0.003673-0.002844
3410.0166730.0087950.0053370.0028990.001391-0.003675-0.002807
3400.0169050.008610.0052060.0029790.001421-0.003677-0.003056
3390.0169150.008460.0051640.0028280.001363-0.004019-0.002883
3380.0171170.0086990.0054360.0028620.001416-0.003645-0.002824
3370.0171560.0088080.005280.0029080.001358-0.003778-0.002962
3360.0175330.0088150.0052570.0030070.001398-0.003711-0.002957
3350.0175480.0087030.0053380.002870.001256-0.003741-0.003031
3340.0177620.0088450.0054280.0030240.001463-0.003912-0.003003
3330.017680.008730.0051340.0029420.001208-0.003943-0.003316
3320.0179310.0087710.0052130.0028870.00106-0.003987-0.003156
3310.0180430.008710.0052770.0029960.001311-0.003953-0.003146
3300.0182430.0086450.0051930.003050.001306-0.004016-0.003097
3290.0184730.0086930.0052520.003020.001497-0.003865-0.002795
3280.0184820.0086250.0050970.0029430.001258-0.003863-0.002948
3270.0186830.008550.0054230.002990.001419-0.003684-0.002834
3260.0185360.0084030.0049470.0028090.001373-0.003688-0.002781
3250.0187510.0087510.0050340.0027050.001485-0.003504-0.002741
3240.0192380.0087680.0050540.0030970.001646-0.003543-0.002839
3230.0193570.0087020.0051570.0028080.001572-0.003345-0.002645
3220.0192550.0086690.0050720.0028440.001574-0.003801-0.002811
3210.0193970.0083990.004850.0025940.001451-0.003549-0.003013
3200.0195880.0086730.0051380.0028380.00147-0.003755-0.002781
3190.0196940.0082210.0051210.0028440.001525-0.003559-0.002763
3180.0199770.0085780.0050810.0029170.001544-0.003703-0.002666
3170.0198570.0086460.0050950.0028780.001588-0.003553-0.002825
3160.0207330.0087770.0055960.0030730.001851-0.003196-0.002486
3150.0206040.0086450.005390.0031510.001822-0.003423-0.002626
3140.0205460.0087380.0052290.0029080.001383-0.003694-0.002903
3130.0208770.0087380.0053430.0029520.001378-0.003381-0.002748
3120.020990.008780.0055280.0032010.001578-0.003578-0.002724
3110.0214440.0088990.005450.0030930.002032-0.003534-0.00267
3100.0214470.0089710.0053260.0029590.001677-0.003774-0.002905
3090.0218140.0092970.0056470.003260.002242-0.00338-0.002497
3080.0219410.00920.0054510.0033490.001852-0.003545-0.002628
3070.0222010.0097720.0059160.0035680.002119-0.00323-0.002452
3060.0223910.0100580.005910.003660.002251-0.003268-0.00221
3050.0227030.0105210.0064870.0040130.002691-0.002988-0.002022
3040.0229550.0112810.0069150.0043950.002905-0.002669-0.001973
3030.0231630.0119390.007650.0047720.003448-0.002524-0.001658
3020.0236650.0128810.0081670.0050580.003537-0.002215-0.001557
3010.0238750.0137210.0091090.0059020.004187-0.001614-0.000927
3000.0243370.0144150.009590.0061710.004582-0.001475-0.000723
2990.0247820.0154420.0104660.0066250.005133-0.001237-0.000427
2980.0252880.0167720.0113120.0074590.005703-0.0007190.00001
2970.0257820.0178550.01220.008090.006202-0.0002580.00054
2960.0265550.0194780.0132950.0090660.0071890.000190.00109
2950.0272460.0210820.0146970.009910.0077270.0010860.001593
2940.027750.0233480.0164850.010970.0090030.0016820.002063
2930.0284530.0261760.0182180.0125630.0102530.0026370.002985
2920.0298070.0288690.0205180.0141070.0116480.0037520.004101
2910.0302720.0312550.0223280.0156420.0129330.0046570.00482
2900.0310130.0332370.0240460.0166980.0138060.0051270.005388
2890.0318220.0356370.0256180.0179330.0148190.0062170.006163
2880.032530.0383770.027820.0193610.0163830.0071890.006906
2870.0335980.0414490.0301980.0211250.0180140.0085180.00824
2860.0343670.0434490.0318650.0223920.0188770.0090520.008833
2850.0353050.0454550.0329090.0235390.0199290.0098180.009454
2840.0359060.0464330.0339450.023950.0205710.0100960.010012
2830.0366770.0474780.0347340.0244270.021020.0106350.010405
2820.0375210.0478940.0350280.0247590.0212710.0108030.010548
2810.0383450.0486640.0354090.025330.0216340.0111050.010885
2800.0393580.0490750.0361030.0256640.0220690.0112920.010982
2790.0404050.0495950.0362020.0257330.0221350.0112390.011325
2780.0414650.049440.0363240.0259460.0224660.0113530.011463
2770.042330.049430.0362820.0257130.0222250.0114230.011523
2760.0427440.0487760.0353630.0251740.0217710.0110670.011047
2750.0433830.0483550.0355060.0253610.021990.0113110.011576
2740.0437040.0474670.0347520.0250250.0219940.0109780.011313
2730.04430.0471740.0345370.0249680.0217850.0110180.011063
2720.0443140.0458180.0333290.0239220.0210780.0103390.010918
2710.0449470.0450440.0326240.0238350.0208510.0102010.010764
2700.0453160.0444470.0324240.0233730.0205870.0100090.010663
2690.0456260.0437270.0318740.0230780.0200910.0097670.01059
2680.0454620.042570.0311080.0223670.0197620.009380.010451
2670.0452520.0414470.030020.0218480.0192570.0090270.010121
2660.0451280.0403110.0295320.0213720.0190650.0086810.009863
2650.0450410.0396510.0286630.0208780.0185130.008530.009433
2640.0450780.038210.0278870.0202080.0180160.0078160.009128
2630.0450130.0368910.026740.0194170.0174070.00740.008864
2620.044750.0359640.0260310.0190440.0169760.0070990.008472
2610.0443580.0353220.0256260.0185870.0167950.0070810.00861
2600.044270.0346550.0250540.0182420.0164490.0068490.008332
2590.0441440.0341890.0247650.0179990.0161980.0063520.008024
2580.0437550.0329990.0237620.0172580.0156170.0059540.007588
2570.0439410.0320310.0231570.0169070.015030.0056670.007234
2560.0434420.0310770.0225150.0162360.0145820.0051280.006796
2550.0433470.0308210.022110.0161280.0143320.005040.007019
2540.0432770.030780.0220890.0161110.0143750.004960.006693
2530.0435470.0310350.0222930.0162630.0146280.0051250.006895
2520.0435810.0309680.0221740.0161080.0144340.0051180.00681
2510.0433150.0306160.0219230.0158130.0141160.0048730.006559
2500.0435140.031010.0222910.0161940.0145120.0051080.006905
2490.0435960.0319380.0230070.0168190.0151010.0054740.007092
2480.0435130.0332110.0238580.0171810.0155660.0057050.007596
2470.0438720.0347520.0251950.0182270.0162560.0064740.00803
2460.0440560.0368010.0269280.019360.017170.0072990.008481
2450.0444310.0397450.0289430.021170.0186830.0082980.009304
2440.0448450.0432610.0316410.0228440.0203140.0094120.01046
2430.0456170.0481890.0354670.0256750.0227330.0111710.012075
2420.0462630.0534690.0397610.0287480.0256510.0133620.014126
2410.0476330.0608230.0453090.0329610.0290950.0160940.016623
2400.048640.0693150.0521670.0378260.0335530.0191720.019287
2390.0501940.0803620.0605940.0440340.0389760.0233210.023041
2380.0521210.0940540.0711340.0515930.0455020.0282990.027662
2370.0538350.1097310.0834240.0603110.0533220.0341850.032731
2360.0563320.1283960.0981150.0709720.062610.0412670.038983
2350.0587790.1496950.1146750.082980.0732960.0489670.046151
2340.0612860.1728040.1326130.0961450.0848730.0577190.053831
2330.0635390.1965710.1512520.1095910.0964510.0666770.061738
2320.0650950.2198490.1694890.1228440.1082840.0751080.069573
2310.0661460.2417670.1866590.1354050.1193910.0835780.077366
2300.0659170.2607270.2016480.1464670.1290470.0907410.084185
WT 1
Fractions
Relative Ratio A280/A500
(D) Wild type rhodopsin
WT
Wavelength (nm)
Absorbance
-
Spin labeling experiment for electron paramagnetic resonance (EPR) spectroscopy Rhodopsin was labeled with 50uM NEM in 50uM NaPi, 0.05%DM in dark for at least 16 hrs Wash with 5mM MES, 0.05%DM 50uM spin label MTSL was added for 3 hrs RT
Rhodopsin was labeled with 50uM NEM in 20mM Tris, 0.05%DM, 150mM NaCl in dark for at least 16 hrsWash with 20mM Tris, 0.05%DM, 150mM NaCl50uM spin label MTSL was added overnight at 4C R9 and R11After extensively washing with 1XPBS, 0.05%DM followed by 2mM NaPi, 0.05% DM of solubilized Rhodopsin-1D4 SepharoseWashing 5mM MES, 0.05%DMLabeled Rhodopsin were eluted 70uM epitope nonapeptide
-
Chart2
-0.002806-0.001931-0.0022550.0012670.0005640.0035270.0045310.0084590.003624
-0.002743-0.001945-0.0021960.0012720.0005050.0035270.0046710.0084940.003611
-0.002748-0.001908-0.0021090.0012720.0006240.003620.0046430.0084520.003583
-0.002803-0.001991-0.0022330.0012310.0005650.0035190.0045840.0083750.003598
-0.002662-0.001826-0.00210.0012630.0006340.0036580.0047040.0084770.003699
-0.002658-0.001877-0.0021190.0012640.0006570.0036540.0047010.0083810.003682
-0.002626-0.001804-0.0020230.0013140.0007210.0037790.0047650.0085580.003774
-0.002695-0.001955-0.0021330.0011630.0006150.003650.0047340.0084290.003655
-0.002586-0.00191-0.0020650.0012640.0006380.0037060.0047720.0084770.003766
-0.002622-0.001901-0.0020880.0012320.0006250.0038220.0047860.0084360.003697
-0.0026-0.001869-0.0021110.0011950.0006250.0038410.0047220.0084190.003735
-0.002591-0.001782-0.0020290.0013380.0007540.0039250.0048570.0084940.00373
-0.002564-0.001838-0.002020.0012880.0007350.0039490.0047840.0084670.003773
-0.002542-0.001893-0.0021120.0012470.000680.0038520.0047660.0083990.003718
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0.0389040.0322240.0215930.0214820.0198040.0235910.02350.0186250.01441
0.0381890.0316680.0211590.0210440.0195410.0234690.0233580.0184780.014314
0.037460.0312940.0208640.0207990.0193570.0233480.0233230.0184290.014319
0.0369260.0308380.0204870.0205920.0192210.0233070.0233230.0184370.014287
0.0363450.0304170.02020.02030.0190140.0231750.0232660.0183460.01425
0.0357740.0300380.0198780.0200930.0188780.0230530.0232650.0182240.014149
0.0351440.0296140.0195570.0197970.0186870.0229880.0231740.0181280.014097
0.0347250.0293270.0193050.01960.0186450.0229260.0232190.0181810.01411
0.034240.0290210.0190490.0194590.0184940.0228450.0232180.018140.014132
0.0338620.0287340.0188270.0191870.0183770.0228380.0231920.0181080.014055
0.03340.0284710.0185970.0190020.0182570.0228290.0232230.0180370.014024
0.0330660.0282360.0183910.01890.0181660.0227580.0232070.0180460.014121
0.032710.0280160.0182340.0187830.0181590.0227510.0232870.0180690.014044
0.0324330.0278650.0180450.018630.0181350.0227130.0233030.018150.01409
0.0320340.0276310.0178580.0184420.0179980.0227930.0232830.0180220.013937
0.0318560.0274640.0177020.0183570.0178920.0226920.0233950.0180720.014064
0.0316250.0273580.0176630.0182720.0180120.0228440.0235110.0180620.014074
0.0313610.0272480.0174580.0181530.0178480.022730.0234730.0180930.014088
0.0312670.0272060.0174550.0181850.0179250.0228490.0236430.018210.014125
0.0310720.027160.0173740.0180430.0179430.0228780.0237230.0182230.014236
0.0309910.0270440.0173080.0180520.0178870.0229140.0238150.0181870.014264
0.0309050.0269880.0171970.0180210.0180160.0229440.0239310.0182910.014292
0.0307630.0269780.0171390.0179080.0179330.0230490.0239760.0182480.014268
0.0307350.026940.0170930.0179630.0179780.0231110.0240590.0183880.014316
0.0306420.0269440.0170560.0178950.017980.0231650.0241640.0182760.014368
0.0305910.0269480.0170620.0178310.0179770.0232080.0242020.0184020.014398
0.0306860.0270520.0171060.0179560.0180660.0233660.0243450.0185320.01452
0.0306890.0270740.0170950.0178510.0182360.0234430.024530.0184620.014514
0.030750.027190.0171170.0179980.0183190.0235370.0246750.018690.014715
0.0307130.0271770.017180.0179960.0183020.0236650.0247230.0186030.014563
0.0307470.0273130.0172050.0180060.0184280.0237580.025030.0186940.014731
0.0308070.0273250.01710.0180860.0185130.0239630.0251690.0188640.014829
0.0310420.0275430.0172910.0182430.018740.0240970.025370.0190120.01495
0.0309890.0276170.0172390.0182270.0187090.0242450.0254840.0191110.014982
0.0311650.0276410.0173150.0182930.018720.0241970.0254460.0190160.014941
0.0313510.027960.0173970.018340.0189930.0244630.0258040.0191940.015188
0.0314950.0280450.0175670.0184510.0190540.0246330.0258530.0193310.015222
0.0317270.0281910.0176340.0185140.019260.0247560.0261120.0193350.015164
0.0323320.0288220.0180750.0188220.019570.0254060.0263050.0195450.015333
0.0324320.0287210.0180250.0189640.0197320.025290.0266370.0198130.015467
0.0325950.0289230.0180610.0189130.0199080.0255140.0267730.0197250.015601
0.0328040.0291530.018130.0191690.0199620.0258020.0269830.0198850.015727
0.03320.0292630.0183030.0191340.020160.0259580.027310.0200830.01592
0.0332220.0295060.0184180.019370.0202480.0261770.0273320.0200750.015966
0.0333840.0296790.0184160.0194080.0203110.0262520.0275520.0202050.015966
0.033820.0300040.018790.0196810.0207770.0265050.0279130.0203760.016193
0.0339590.0300690.0186230.0196250.020660.0265580.0279040.0205030.016258
0.0342130.0303480.0188330.0198710.0208410.0267280.0282290.0205870.016292
0.0346040.0305960.0189980.019980.0210360.0269290.0284410.0208480.016487
0.0348790.0307290.0191180.0202880.0212070.0271740.0286350.0209280.016576
0.0351250.031030.0192490.0203130.021380.0273180.0288260.0209280.016677
0.0355810.0314210.0195490.0205420.0216490.0276760.0292110.0211820.016935
0.0358520.0316740.0196950.0207030.0218010.0278040.0293380.0213840.017065
0.0361450.0317970.0197020.0207060.0219760.0279410.0295490.0213810.017092
0.0364060.0319660.0198650.0208950.0220390.0280720.0297630.0215450.017154
0.0367610.0323020.0199150.0211220.0224040.028380.030.0216960.017294
0.0372020.0325790.0201190.0213110.0226140.0286650.0302950.0219420.017613
0.0374790.0328210.0204670.0215840.0227860.0289160.0305940.0221130.017783
0.0376890.0331170.020480.0216880.0229770.0291260.0308570.0222230.017856
0.0383010.033390.0207580.0218550.0232410.0294480.0311290.0225010.018047
0.0385330.0337020.0208420.0220510.0233510.0295350.0313720.0225760.018024
0.0389080.0339840.0209530.0222440.0236060.0298560.0315950.0226570.018221
0.0392650.0342950.0212030.0223480.0237610.0300240.0318470.0228320.018303
0.0396560.0345240.0212820.0224980.0240240.0303070.0320690.0229260.018421
0.0399950.0347480.0215280.0227250.024190.0305750.0323440.0231480.018544
0.0402810.0350090.0216560.0228530.0244060.0308060.0324930.023220.018665
0.0405920.0352520.0217870.023020.0246770.031020.0328280.0234850.01881
0.0409130.0356470.0218520.0231680.024820.0313430.033220.0235760.018995
0.0412950.0357810.0220860.0233460.0251230.0315170.0333640.0237710.019075
0.0415250.0359790.0221180.0235020.0253110.0318670.0337640.0238750.019267
0.0417890.0361210.0221260.0236280.0254850.0321250.0339880.0240940.019339
0.0419840.0362620.0223350.0237060.0256310.0322520.0341160.0242850.01949
0.0418930.036420.0222550.023750.0256920.0323890.0342820.0242470.01961
0.0421630.036540.0223670.0239040.0259680.0327880.0345950.0243970.019674
0.0423090.0365420.0221650.0238260.0259430.0328380.0346830.0243660.019796
0.0423530.0365630.0222070.0237630.0260050.0329030.034930.0245860.019826
0.0424310.0368230.0223540.0240080.0262740.0333270.0351560.0247290.020026
0.0425370.0367420.022370.0240510.0264020.0333850.0355970.0249080.020193
0.0426550.0367710.0222930.0240220.0266180.0337130.0358640.0250540.020321
0.0426080.0367530.0222330.024220.0266880.0337570.0359510.0251650.020367
0.0426690.0369480.0221990.0241140.0268480.0340180.0361940.0251740.020552
0.0427770.0368830.0222120.0242530.0269680.0340470.0364310.0254790.020683
0.0428740.0369770.0222120.0242740.0271150.0344840.0367540.0256090.020667
0.0428210.037030.0221720.0242340.0271380.034710.0369760.0257460.020929
0.0428270.0369530.0220650.0242720.0271360.0347230.0372620.0256860.020914
0.0431280.0369980.0220.0243360.0274660.0350330.0375950.0259840.021126
0.043020.0370510.021910.0243390.0275320.0351750.0377920.026190.021268
0.0427750.0369660.0217890.0243270.0275840.035280.0380110.0261580.021367
0.0428380.0368930.0218010.0241640.027750.0354620.0381840.0262340.021405
0.0426890.0368510.0215380.0239990.0276940.0354940.0383730.0263810.021394
0.0427220.0367290.0215510.0241620.0279150.0357450.0385770.026570.021556
0.0425050.0366880.0214350.0240370.0278910.0358730.0385520.0263780.021481
0.042570.0369410.0214760.0242890.028180.0362320.0391380.0268540.021918
0.0426080.0368990.0215020.0242910.0282950.036190.0394570.0268330.021924
0.0432140.0372630.0212870.0248640.028840.0371150.0399670.0273570.022636
0.0431480.0373640.0210180.024910.0293090.0373530.040190.027160.022609
0.043270.0373320.0212010.0248360.0292120.0376120.0404510.0274370.022976
0.0435560.0374890.0214330.0251260.0294310.0380650.0412410.0278150.023424
0.0427850.0371010.0207940.0245170.0292060.0377630.0407970.0276510.023164
0.0434470.0376370.0213180.025310.0299250.0384950.0414950.0279450.023371
0.0431450.0372520.0208840.0249250.0298190.0385820.0414330.0277450.023166
0.0432320.0374650.0209940.0249530.0298730.0387370.0418350.0282190.02343
0.0434040.0375020.0209330.0249550.0302820.039050.0421510.0281640.023303
0.0433820.0375110.0208130.0249220.0303110.0393190.0423110.0282860.023679
0.0436720.037830.0210.0251860.0302770.0396980.042670.0283560.023795
0.0436690.0378730.0209570.0252980.0306720.0399920.0431650.0286090.024091
0.0436910.0379110.021070.0254590.0307150.040030.0433320.028750.024333
0.0441760.038350.0213250.0255450.0314040.0409540.0439130.0293280.024532
0.0442520.0385030.0211220.0256440.0312870.0410930.0444130.0293150.024842
0.04460.0386690.020960.0256730.03160.0413360.0444010.0294440.024716
0.0448820.0388450.0212690.0260790.0321280.0416580.0453930.0298020.025054
0.045590.0394240.0216450.0264560.0323970.0421080.0457190.030230.025543
0.0462970.0401490.0221380.026860.0327440.0426170.0463190.030570.025617
0.0472130.0411170.0226530.02740.0333940.043140.0468940.0307850.026058
0.0486720.0422220.0234540.0279010.0339820.0438470.0475110.0313430.026495
0.0506870.0437180.0245250.0289230.0347880.0448310.0484080.0319890.027131
0.0532870.0458480.0260330.0302150.0362910.0458260.0494260.0326720.027669
0.0567980.0486460.027870.0318510.0374160.0472810.050550.0334310.028196
0.0604280.0517220.0302320.033490.0391660.0486340.0518810.0344390.029032
0.0645860.0549140.0324170.0354810.0405340.0498990.0529520.0353230.02993
0.0685840.0584840.0348540.0374330.0424450.0514240.0545050.0364610.030725
0.0728040.0616980.0372910.0394220.0441080.0531180.0557530.0374120.031418
0.0772520.0653250.0398930.0415410.0457220.054790.057220.0384750.032475
0.0819660.0690510.0425970.0437180.0474670.0563660.0588280.0395860.033385
0.0872380.0731950.0457160.0461810.0497280.0581180.0605340.0407060.034557
0.0936870.0788790.0495090.0492430.0522130.0604540.062580.0425050.035727
0.1023080.0857710.0544090.0533690.0557770.0632070.0649310.0440110.037445
0.1131780.0944120.0609370.0585650.0595350.0667950.0680050.0465140.039183
0.1257530.1046560.0682960.0644370.0646790.0708710.0712440.0491640.041502
0.1374460.1140140.0750790.0695870.0688040.0743850.0742410.0513270.04335
0.1477380.1225880.081570.074770.0730680.0780310.0774080.0541320.045486
0.1577040.1303860.0871770.079420.0767040.0814240.0803040.0560980.047175
0.1692160.13970.0938430.0848230.0811090.0851540.083310.058550.049293
0.1816140.1497040.1014050.0907580.0858370.0889120.0867110.061230.051486
0.1941130.1599020.1086270.0965550.0904960.0931460.0902740.0637930.053668
0.2048160.16860.1149490.1014310.0948290.0965980.0930370.0659580.055604
0.2122880.1742410.119070.1049210.0974260.0991390.0954590.0676890.057167
0.2170930.1786240.1221680.1075150.0999920.1015290.0974270.069230.058152
0.2205420.1813210.1241750.1092140.1014570.1028760.0988470.0701140.05928
0.2239280.1843740.1260620.1108690.1032810.1047310.1004940.0712170.0602
0.2274250.187110.1279480.1123280.1047460.1063060.1018380.0723190.06101
0.2303070.1895820.1294420.1137950.106390.1083190.1040490.0734620.062084
0.2313750.1908050.1299670.1142720.1073430.1092450.1053930.0744330.062986
0.2317880.1910690.13020.1148710.1081170.110310.1070860.0750410.063697
0.2306270.1904280.1291130.1142780.1084390.1112570.1080330.0756310.064452
0.2281610.1885280.1275230.1129360.1079490.1111740.108530.0753390.064393
0.2249840.1861510.1257940.1116830.1073410.1110150.1086790.0753850.064294
0.2213490.183220.1236250.1099930.1064350.1105780.1085390.074920.06412
0.2168220.179760.1207780.1079920.1050270.1095130.107860.074520.063832
0.2126890.1766810.1183840.1061230.1039140.1088130.1076320.0739580.063432
0.2091450.1736670.1160040.104330.1028580.108520.1077070.0736540.063316
0.2059640.1715410.1141990.1031010.1023670.1083770.1079770.0735680.06331
0.2027360.1692240.1120860.101950.1017830.1082520.1085140.0733320.063133
0.1980660.1651970.1089850.0992620.1003410.1074980.1079030.0728030.062819
0.1923370.1610350.105740.096640.0989190.1068150.1079920.0726620.062849
0.188540.1579230.1030710.0950490.0979030.1061220.1076270.0720410.062534
0.1856620.1557680.1014080.0937280.0971250.1059740.1076590.0718680.062585
0.1796150.1508710.0978660.0907760.0951890.1045360.106550.0709120.061437
0.1738250.1464180.0943820.088330.0937370.103460.1061050.0702440.061263
0.1704860.143870.0921010.0868770.0927430.1033130.106340.0700480.06098
0.1678780.1422790.0905720.0858450.0925890.1032520.1065670.0699150.061262
0.1665890.1409330.0895730.085150.0923140.1036170.1070980.0699490.061376
0.1648360.139790.0884480.0845930.0922130.1037360.1076240.0701460.061488
0.1614410.1371270.0863630.082940.0915810.1037040.107840.0702950.06174
0.1570330.1335950.0835540.0808890.0906250.1031170.1079720.0698670.061497
0.1548030.1320650.0821340.0799720.0902740.1034990.1084730.0699930.061515
0.1543560.1319050.0816170.0795910.0906270.1042620.109590.0703730.061969
0.1554480.133030.0820150.0801910.0916070.1055430.1108850.0711690.062714
0.1571370.1342780.0828550.0811220.0928120.1070010.1125450.072170.063768
0.1580810.1354510.0834040.0817560.0939380.1084690.1142170.0729650.064656
0.1598780.1368150.0840350.0825080.0952820.1100270.1157880.0737850.065581
0.1638120.140330.0861010.0844570.0974710.1126530.1186280.0754390.066779
0.1694230.1453970.089260.0873160.100410.1155340.1214840.0773940.068651
0.1775890.1519620.0939440.0912020.1042690.1193390.1251530.0800050.070636
0.1870050.1600230.0993830.0958180.1086870.1237910.1294470.0827610.073393
0.1991740.1699880.10610.1015820.114040.1287280.1341650.0860060.076269
0.2142850.1825110.1149670.1087180.1206980.1350440.1401330.0900090.079658
0.2332610.1981280.1256670.117640.1285520.1422260.1467130.0946510.0837
0.257240.2180330.1398310.1291880.1386090.151480.1551760.1006390.088789
0.2868140.2420390.1569450.1431420.1508410.1625590.1650640.1079650.095035
0.3225560.2715230.177830.1600730.1654540.1756540.176730.116270.1024
0.3670470.3081270.2038040.1813150.1834280.1917940.1911930.1268120.111295
0.4223220.3535080.236260.2073890.2059170.2118350.2090970.139810.122396
0.4899590.4091040.275910.2396440.2333340.2362240.2307550.1558350.136
0.5694690.4743030.3223310.2772440.2652380.26440.2559780.1741430.151759
0.660930.549780.3768070.3209160.3019070.2969530.2850350.1954590.170124
0.7634630.6338750.437490.3699430.343220.3332420.3174090.2194580.190673
0.873120.7240560.5034350.4221270.3865730.3711440.3513820.2445160.212198
0.9849780.8156490.571630.4759910.4306490.4088390.3852930.2693270.233885
1.0930310.9056470.6388010.5285210.4728360.4449070.4172110.2934590.254529
1.1918880.9867620.6999360.5756530.5111660.4762650.4447350.3146060.272907
1.2752821.0558340.7524570.6163260.5431610.5025280.4670930.3323660.287737
Elute 1
Elute 2
Elute3
Elute 4
Elute 5
Elute 6
Elute 7
Elute 8
Elute 9
Wavelength (nm)
Absorbance
Eluted R11 with spin labeling
R11 1
650-0.003048-0.002806-0.001931-0.0022550.0012670.0005640.0035270.0045310.0084590.0036240.0097270.1204480.0946460.0697460.0559950.0443580.037890.0310.0226370.020102
649-0.00308-0.002743-0.001945-0.0021960.0012720.0005050.0035270.0046710.0084940.0036110.01006241380.12460137930.09790965520.07215103450.05792586210.04588758620.03919655170.03206896550.02341758620.0207951724
648-0.00298-0.002748-0.001908-0.0021090.0012720.0006240.003620.0046430.0084520.003583totalper plate
647-0.003104-0.002803-0.001991-0.0022330.0012310.0005650.0035190.0045840.0083750.0035980.52401620690.0131004052
646-0.00304-0.002662-0.001826-0.00210.0012630.0006340.0036580.0047040.0084770.003699
645-0.002986-0.002658-0.001877-0.0021190.0012640.0006570.0036540.0047010.0083810.003682280-6500.0369530.2331130.1915130.1316970.1125280.1058260.1047920.0995180.0650030.05846
644-0.002977-0.002626-0.001804-0.0020230.0013140.0007210.0037790.0047650.0085580.003774500-6500.0097270.1204480.0946460.0697460.0559950.0443580.037890.0310.0226370.020102
643-0.002977-0.002695-0.001955-0.0021330.0011630.0006150.003650.0047340.0084290.003655280/5003.79901305641.93538290382.02346639061.88823731832.00960800072.38572523562.76569015573.21025806452.87153774792.9081683415
642-0.003042-0.002586-0.00191-0.0020650.0012640.0006380.0037060.0047720.0084770.003766
641-0.003037-0.002622-0.001901-0.0020880.0012320.0006250.0038220.0047860.0084360.003697stdev0.6392072302
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4540.0053560.0748970.0595680.0424040.0377610.0311550.0316930.0287050.0240310.018588
4530.0052810.0736190.0585620.0417190.0371220.0307880.0314010.0285110.0237880.018386
4520.0052870.0722280.0575320.0408530.0364910.0303560.0311560.0282770.023610.018199
4510.0052180.0709720.0564080.0401840.0359030.0298090.0307560.0280280.0232720.017961
4500.0052060.0696210.0555180.0393860.0353360.029540.0304210.0278510.0231440.017822
4490.0052320.0684290.0546320.0386460.0348060.0291770.0302650.0276890.0229580.01773
4480.0051060.0670330.0535390.0379330.0341840.0286560.029980.0275310.0227180.017619
4470.0049050.0655740.0523520.0370050.0335370.0280970.0295580.0270790.0224660.01733
4460.0049750.0644140.0514380.0362690.0329730.0277390.0293320.0270030.0223140.017266
4450.0048210.0629960.0503550.0355090.0322260.0273390.0288960.0266690.0220140.01699
4440.0047520.0617780.0493970.0348210.0317710.0269520.0286050.026610.0219630.016882
4430.0047030.0604080.0483860.0340060.0311530.0264080.0283380.0263680.0216550.016752
4420.004720.0591670.0474460.0332530.0305760.0260670.0280620.0261520.0214290.016589
4410.0046730.058230.0466540.0326620.0301810.0257350.0279020.0260280.0214840.016588
4400.0045240.0567390.0455770.0318980.0294810.025270.0274820.0257940.0211290.016203
4390.0044680.0557460.0449320.0311860.0289750.0250030.0271640.0255780.0210280.016126
4380.0044450.0545890.0440780.0305990.0284760.0245640.0269610.025390.0207860.016068
4370.0043610.0534060.0430380.0299290.0281180.0242870.0267750.0253350.0205840.015914
4360.0043050.0522760.0423130.0293590.0276870.0240140.0265470.0252180.0204020.015871
4350.0041820.051150.0414350.0288370.0270390.0236870.026350.0250570.0204050.015877
4340.004280.0501810.0406030.0280780.0266630.0232770.0261210.0248020.0202410.015579
4330.0042010.0489420.0398030.0273360.0261090.0228710.0257650.0246740.0199570.015548
4320.0043260.0481360.0391910.0268870.0256860.0226570.02560.0246160.0199640.015455
4310.004140.0470750.0382820.0262560.0252390.0223770.0254260.0244310.0197190.015255
4300.0040930.0461310.0376320.0257480.0248990.0221290.0250860.0244340.0197220.015232
4290.0040080.0452210.0368750.0251790.0242490.0216180.0248650.0241030.0194570.015027
4280.0040120.0444150.0361690.0247460.0239780.021550.0248620.0242050.0193540.015129
4270.0039210.0434510.0356050.0241740.0235390.0211020.0245680.0239720.0192470.014884
4260.0038990.0426650.0348460.0237180.0230730.0208240.0243440.0237740.0190440.014692
4250.0040330.0418790.03440.023350.0228510.0207980.0242320.0237930.0190890.014731
4240.0039440.0410170.0337710.0228020.0223140.0203020.0241020.0237490.0188830.014678
4230.0038290.0402630.0332290.0223070.0221010.0201450.0239290.0236110.0188110.014641
4220.003720.0394810.0326530.0219610.0216590.019910.0237190.0234010.0187010.014495
4210.0038590.0389040.0322240.0215930.0214820.0198040.0235910.02350.0186250.01441
4200.0037340.0381890.0316680.0211590.0210440.0195410.0234690.0233580.0184780.014314
4190.0038120.037460.0312940.0208640.0207990.0193570.0233480.0233230.0184290.014319
4180.0038410.0369260.0308380.0204870.0205920.0192210.0233070.0233230.0184370.014287
4170.0038080.0363450.0304170.02020.02030.0190140.0231750.0232660.0183460.01425
4160.0037610.0357740.0300380.0198780.0200930.0188780.0230530.0232650.0182240.014149
4150.0037320.0351440.0296140.0195570.0197970.0186870.0229880.0231740.0181280.014097
4140.0038490.0347250.0293270.0193050.01960.0186450.0229260.0232190.0181810.01411
4130.0037770.034240.0290210.0190490.0194590.0184940.0228450.0232180.018140.014132
4120.0038120.0338620.0287340.0188270.0191870.0183770.0228380.0231920.0181080.014055
4110.0037790.03340.0284710.0185970.0190020.0182570.0228290.0232230.0180370.014024
4100.0038520.0330660.0282360.0183910.01890.0181660.0227580.0232070.0180460.014121
4090.003920.032710.0280160.0182340.0187830.0181590.0227510.0232870.0180690.014044
4080.0038820.0324330.0278650.0180450.018630.0181350.0227130.0233030.018150.01409
4070.0038350.0320340.0276310.0178580.0184420.0179980.0227930.0232830.0180220.013937
4060.003880.0318560.0274640.0177020.0183570.0178920.0226920.0233950.0180720.014064
4050.0038750.0316250.0273580.0176630.0182720.0180120.0228440.0235110.0180620.014074
4040.0038860.0313610.0272480.0174580.0181530.0178480.022730.0234730.0180930.014088
4030.0040650.0312670.0272060.0174550.0181850.0179250.0228490.0236430.018210.014125
4020.0040270.0310720.027160.0173740.0180430.0179430.0228780.0237230.0182230.014236
4010.0041010.0309910.0270440.0173080.0180520.0178870.0229140.0238150.0181870.014264
4000.0041210.0309050.0269880.0171970.0180210.0180160.0229440.0239310.0182910.014292
3990.0041130.0307630.0269780.0171390.0179080.0179330.0230490.0239760.0182480.014268
3980.0041230.0307350.026940.0170930.0179630.0179780.0231110.0240590.0183880.014316
3970.0042020.0306420.0269440.0170560.0178950.017980.0231650.0241640.0182760.014368
3960.0042470.0305910.0269480.0170620.0178310.017
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