Structural Genomicsof Mycobacteria
Pedro M. Alzari
Why?Protein fold catalogBiomedical interestFunction discovery
How?High-throughput methods
Structural Genomics
The Structural Genomics Pipeline
Gene cloning
Protein purification
Crystallization
NMR,X-ray Data collection
3D structure determination
Protein expression
Analysis (annotation)
Flow optimizationWeb DatabasesAutomation
Miniaturization
Estimates of TB burden (1997)
New cases in 1997
New cases : 7,962,000 (1/4 sec)TB deaths : 1,871,000 (1/17 sec)
Infection prevalence : 1,855,880,000 (32%) Dye et al, JAMA,1999
Isoniazid, Rifampicin,Pyranizamide, Ethambutol
No new anti-TB drugin more than 40 years
M. hiberniaeM. terrae
M. nonchromogenicumMCRO6
M. cookiiM. celatum
M. shimoideiM. asiaticum
M. gordonaeM. marinumM. ulcerans
M. tuberculosis ComplexM. lepraeM. szulgai
M. malmoenseM. haemophilum
M. gasti / kansasli
M. « paraffinicum »M. scrofulaceum
M. intracellulareM. paratuberculosis
M. avium
M. intermediumM. interjectum
M. genavenseM. simiaeM. triviale
M. peregrinumM. fortuitum
MCRO19MCRO18
M. xenopi
Phylogenetic tree (16S rRNA gene seqs)Phylogenetic tree Phylogenetic tree (16S (16S rRNA gene seqsrRNA gene seqs))
Slowly growing mycobacteria
M. tuberculosisM. africanumM. canettiiM. microtiM. bovisM. bovis BCG
Tuberculosis
Leprosy
M. smegmatis
• Target selection
• The pipeline
• Results
Structural Genomics of Mycobacteria
Actinomycetes-restricted genesM. tuberculosis
(3959 genes)
M. leprae(reductive evolution)
(1604 genes)
Actinomycetes [ other than mycobacteria ] 470
218
114
136
Mt-specific
Actino-specific
Myco-specific
Ml-specific
267solubleproteins(predicted)
Gene essentiality in TB largely confirmed by high density mutagenesis
Sassetti et al, Mol.Microbiol., 2003
EXTRACELLULARREPLICATIVE
LIFE
Granulome undergoes caseation
Pulmonary cavity
Adapted from, Nature Rev. Mol. Cell Biol., 2, 569-586 (2001)
Pulmonary cavity
Aerosol spread of infectious
INTRACELLULARREPLICATIVE
LIFE
Infected alveolar macrophages
Granuloma or tubercle
DORMANCE
Recruitment of mononuclear cells
Blood vessel
REACTIVATION(old age, malnutrition,
HIV-co-infection)
Life cycle of M. tuberculosis
S.coelicolor0
1
2
3
4
5
6
7
8
9
10
P.aerug. E.coli M.tb B.subtilis V.cholerae Synechoc. M.leprae0
10
20
30
40
50
60
70
80
90Histidine KinasesResponse regulators
Geno
me
size
(M
B)
Num
ber
of g
enes
Eukaryotic-like signaling elements
Ser/Thr protein kinases
Two-component systems
RD5 (mpt40)Rv 2345
plcC plcB plcA PPE PPE
IS61102625 2626 2627 2628 2629 2630 2631 2632 2633 2634 2635 26362624 kb
Rv 2346/47/48
ephA Rv3618 lpqG
Rv3616 Rv3619/20 PPE PE
RD8
4061 4062 406340604059405840574056 kb
RD10Rv0221 echA1
Rv0223
0265 0266 0267 02680264
Rv0224
kb
RD9
Rv2075/76
Rv2074
Rv2073cobLcobM
2330 23332329 23322331 kb
RD12
3483 3484 3485 3486 3487 3488 3489moeB
3490 kbcys3A
sseCmoaE
Rv3120/21/22/23 Rv3124 RD13
1401 1402 1403 1404 1405 1406deaD
1407 kbRv1254
Rv1255
Rv1256 Rv1257 Rv1258RD11 (phiRv2)
Rv2645/46/47 IS6110
Rv2650/51 Rv2652/53/54Rv2655/56/57/58/59/60/61
glyTcysTvalU
valTRv2644
2970 2971 2972 2973 2974 2675 2976 2977 2978 2979 2980 2981 kb2969
RD3 (phiRv1)
REP ’
Rv1573/74/75
Rv1576/77/78Rv1579/80/81Rv1582/83/84/85 / 86 REP
bioB17801779 1781 1782 1783 1784 1785 1786 1787 1788 1789 1790 kb
bioD 1778
Rv1571
oriC
Mycobacterium bovis BCG Pasteur 1173 P2
PPEPPEPPE
Rv3424alr IS1532
RD63841 38443842 3843 3845 3846 3847 3848kb
Rv3430
RD1PE/PPE
4359 4360 4361 kb4351 43544352 4353 4355 4356 4357 43584349 4350Rv3871 Rv3874esat6 Rv3876 Rv3878
Rv3879 Rv3880/81
Rv3877
Rv1771200019991998 2001 2002 2003 2004 2005 2006 2007 2008 2009 kb
Rv1766/67
IS9’Rv1765
Rv1770Rv1769PE_PGRS
Rv1772 Rv1774
Rv1773
RD14
RD4gmdA epiA Rv1513
Rv1514/15 Rv1516
Rv1517
Rv1508
Rv1509Rv151017001699169816971696 1701 1702 1703 1704 1705 1706 1707 1708 1709 kb
Rv1505/06/07
kb
RD7 RD2Rv1965Rv1964 mce3 Rv1967Rv1968Rv1969 lprM Rv1971/72/73/74/75
Rv1976
Rv1977 Rv1978
Rv1979 mpt64 nrdFRv1982
PE-PGRS
Rv1984 Rv1985
Rv1986Rv1987/882208 22092207 22202219221822172216221522142213221222112210 2221 2222 2223 2224 2225 2226 2227 2228 2229 2230 2231 2232
Rv1963 Rv1989
Virulence factorsWT +RD1
BCG Pasteur
M. microtiOV254
Time after infection
CFU
/Lun
gs
RD1 recombinants inM. bovis BCG and M. microti
ESAT-6expression
Mouse immunogenicity
Brodin et al, Infect Immun, 2002; Pym et al, Mol Microbiol, 2002; Nature Med, 2003.
?
EsxA (ESAT-6)
EsxB (CFP-10)EsxAB
EsxAB
Mycolic acids
Cytoplasmicmembrane
Peptidoglycan
Arabinogalactan
R.Brosch, IPR.Brosch, IPSystematic gene knock-out (secretion machinery)Systematic gene knock-out (secretion machinery)Virulence of EsxA mutants (host-parasite interactions)Virulence of EsxA mutants (host-parasite interactions)
• Target selection
• The pipeline
• Results
Structural Genomics of Mycobacteria
Crystallization
Cartesian Technologies nano-dispenser
103No hints orsalt crystals
19Exploitable hints
28Diffracting
crystals
2004
Parallel multi-microfermentors (MMF)
1
10
100
0 4 8time
OD140 mg
- +
M.tb AMPK
J.Bellalou, P.Beguin, IPJ.Bellalou, P.Beguin, IP
Web Databases
http://www.pasteur.fr/SGMF. Guillemot, IPF. Guillemot, IP
• Target selection
• The pipeline
• Results
Structural Genomics of Mycobacteria
0102030405060708090
100
Processedtargets
Cloned Soluble proteins Purified Crystals Structures
TB consortium (central facilities) Institut Pasteur
735 426
The solubility bottleneck(sept 2004)
16 10
356
115
65
Rv0049 ML2689 Rv0483 ML2446 Rv1332 ML1166
Rv0098 ML1993 Rv0504 ML2425 Rv1361 ML1182
Rv0116 ML2664 Rv0546 ML2261 Rv1446 ML0580
Rv0146 ML2640 Rv0635 ML1910 Rv1794 ML1540
Rv0177 ML2597 Rv0636 ML1909 Rv1828 ML2075
Rv0184 ML2604 Rv0637 ML1908 Rv1830 ML2073
Rv0185 ML2605 Rv0819 ML2193 Rv1846 ML2063
Rv0201 ML2616 Rv0867 ML2151 Rv1883 ML2031
Rv0216 ML2627 Rv0885 ML2135 Rv1884 ML2030
Rv0288 ML2531 Rv0910 ML2113 Rv1891 ML2023
Rv0289 ML2530 Rv0966 ML0169 Rv1906 ML2010
Rv0292 ML2527 Rv1094 ML1952 Rv1919 ML1983
Rv0313 ML2518 Rv1109 ML1939 Rv1976 ML1791
Rv0356 ML0279 Rv1155 ML1508 Rv2054 ML1444
Rv0358 ML0281 Rv1182 ML1230 Rv2525 ML1190
Rv0455 ML2380 Rv1222 ML1077 Rv3020 ML2532
Rv0464 ML2465 Rv1252 ML1099 Rv3867 ML0056
Rv0466 ML2463 Rv1259 ML1105 Rv3873 ML0051
Rv0477 ML2452 Rv1277 ML1119 Rv3876 ML0048
M.tb M.leprae M.tb M.leprae M.tb M.leprae
cloned no expr. insoluble solubleOrthologous genes
0
10
20
30
40
50
60
genes TB ML TB or ML
37% 35%
56%
Soluble proteins
Optimizing protein expression parametersOptimizing protein expression parameters
PCRPCR In vitroIn vitroexpressionexpression
CloningCloning
Testing protein expression before bacterial cloning
J.M.Betton, IPJ.M.Betton, IP
T7 promoter T7 terminator gene
ccdBori
bla
cat
aphpCR
E.coli(BL21)
Bi-directional cloning of optimallyexpressed PCR fragments
SrfI SmaI
RNA polymerase T7 E.coli lysate (S30)
geneprotein
Cell-free transcription/translation
Parameter optimizationParameter optimization
PCR:
Optimizing for mRNA secondary structureOptimizing for mRNA secondary structure
2 3 8765 109WT 1 4600
500
400
300
200
Band size(bp)
2 3 8765 109WT 1 4
1000
800700600500
400
300
Band size(bp)
2 3 8765 109WT 1 4
32.5
25
16.5
6.5
Protein MW(kDa)
Gene amplification
T7 regulation sequences
RTS expression(anti-His6 Abs)
Met Pro Thr Tyr Ser Tyr GluWT ATG CCG ACC TAC AGC TAC GAG
M1 ATG CCA ACT TAT TCA TAT GAAM2 ATG CCA ACT TAT TCA TAT GAGM3 ATG CCA ACA TAT TCA TAT GAGM4 ATG CCA ACC TAT TCA TAT GAAM5 ATG CCA ACT TAC TCA TAT GAAM6 ATG CCA ACT TAC TCT TAT GAAM7 ATG CCA ACC TAT TCA TAT GAGM8 ATG CCA ACT TAT TCA TAC GAGM9 ATG CCA ACT TAT TCA TAC GAAM10 ATG CCA ACA TAT TCA TAC GAG
ML0180ML0180WT WT M3M3
Total Soluble
BL21 expressionBL21 expression
Silent mutations (ProteoExpert)
RBSSTARTRBS START
X
Optimizing the Optimizing the expression of expression of solublesolubledomainsdomains
kinase domain
Deletions
Expressionclone
1
2
3
5
4
6
7
8
TM
kDa
ImmunoBlot (anti-His6 antibodies)
1 4 5 6 7 832 9
GFP
100 - 75 -
45 -
30 -
20 -
In vitro expression
Rv0015cRv0015c
transcriptionDNA librarymRNA
translation
selectionmRNA elution
reversetranscription
and PCR
mRNA-ribosome-folded proteincomplexes
Immobilized ligand
mRNA
DNA
(± introductionof errors)
= mutation
In vitro evolution (ribosome display)
in vitro
library of variants ofthe target protein(s)
Esx-1 system
immobilizedligand
sélection
= mutation
TargetTarget
proteinprotein
foldingfoldingreporterreporter
Selection for solubility
F. Pecorari, IPF. Pecorari, IP
Rv0014c279 Rv0018cRv0014c331 Rv0813c Rv0877Rv0733 Rv1846c
Rv1908c Rv2238 Rv2461cRv2428Rv2276 Rv2543 Rv2610c
Rv2667 Rv2714 Rv2883c Rv2991 Rv3628 ML2640
Rv3849
Rv1155
Rv2171 Rv3013Rv1399c Rv2945cRv1208 Rv2125
From structure to function:
structure-based
functional annotation
Structural Genomics of Mycobacteria
. . : *. : : . : * **** : . ***: * ****** *. : * . ************* *. . : *: * ***** . : *** : **. : *: * *: * * . **: ****: |15607953|ref|NP_215328.1| - - - - - - - - - - MSSGAGSDATGAGG- - VHAAGSGDRAVAAAVERAKATAARNI PAFDDLPVPADTANLREGADLNNALLALLPLVGVWRGEGEGRGPD- GDYRFGQQI VVSHDGGDYLNWESRSWRLTATGDYQEPGLREAGFWRFVADPY 137|41406741|ref|NP_959577.1| - - - MVCALHAVPAHHRRVVTPAGDDPSGPAGSGDRAVAAAAERAKLTAGRNI PSFDDLPLPADTANLREGANLSDALLALLPLVGVWRGEGEGRGHD- GDYRFGQQI VVSHDGGDYLNWEARSWRLNDTGDYQERGLRETGFWRFVRDPD 146|15828179|ref|NP_302442.1| - - - - - - - - - - MTSDEVRDGAGSPADSSKGNKCTAAGMFQAAKRSTVSAARNI PAFDDLPVPSDTANLREGANLNSTLLALLPLVGVWRGEGEGRGPN- GDYHFGQQI VVSHDGGNYLNWEARSWRLNDAGEYQETSLRETGFWRFVSDPY 139|25029027|ref|NP_739081.1| MSENETSKTGGNAGVPGSGADAPSLSDSPAI SGNDAVNLAAEQAKNTAHRNI PTLDDLPI PEDTANLRLGPNLHDGLLALLPLVGVWRGEGQADTPEGGQYSFGQQI I FSHDGENYLSFESRVWRLDSEGGTVGPDQRETGFWRI N- - - - 146|19553775|ref|NP_601777.1| MSENSTPN- - - NPVVPGAGADGPSLSDSASI SGSDAVNLAAEQSKSTAHRNI PGLGDLPI PDDTANLREGPNLHDGLLALLPLVGVWRGEGQADTAEDGQYAFGQQI TFAHDGENYLSFESRMWKLDEEGNPTGVDQRESGFWRI N- - - - 143|38234482|ref|NP_940249.1| MPSK- - - - - - - QVGWWTMSEHTNDEVQPTALSGNDAVNRAAEQWKESAHRNI PGLGDLPI PDDTANLREGPNLHDGLLALLPLVGVWRGTGHADTAEEGQYAFGQQI TFAHDGENYLTYESRI WKLDDEGNSTDLDYRESGFWRI S- - - - 139 ruler 1. . . . . . . 10. . . . . . . . 20. . . . . . . . 30. . . . . . . . 40. . . . . . . . 50. . . . . . . . 60. . . . . . . . 70. . . . . . . . 80. . . . . . . . 90. . . . . . . 100. . . . . . . 110. . . . . . . 120. . . . . . . 130. . . . . . . 140. . . . . . . 150
. : **. : : *: * . *: : **. * . : *: : : : : * : * . *****: : . : *. : *: **: . . : *. : **. * *. *|15607953|ref|NP_215328.1| DPSESQAI ELLLAHSAGYVELFYGRPRTQSSWELVTDALARSRSG- VLVGGAKRLYGI VEGGDLAYVEERVDADGGLVPHLSARLSRFVG 226|41406741|ref|NP_959577.1| DPSESQAI ELLLAHSAGYVELFYGRPRTQSSWELVTDALARSRSG- VLVGGAKRLYGI VEGGDLAYVEERVDADGGLVPHLSARLSRFAG 235|15828179|ref|NP_302442.1| DPTESQAI ELLLAHSAGYVELFYGRPRNASSWELVTDALACSKSG- VLVGGAKRLYGI VEGGDLAYVEERVDADGGLVPNLSARLYRFAG 228|25029027|ref|NP_739081.1| - - - LQDEI EFVCAHASGVVEI YYGQPI NERAWELESASTMVTATGPVTLGPGKRLYGLLPTNELGWVDERL- VDKELKPRMSAQLHRI I G 232|19553775|ref|NP_601777.1| - - - LKDEI EFVCTHAGGVVEI YYGQPLNERAWQLESASTMVTATGPSTLGPGKRLYGLLPTNELGWVDERL- VGDALKPRMSAQLTRVI G 229|38234482|ref|NP_940249.1| - - - LKDEI EVVLTHSTGVAEI FYGEPMNERAWQI ESASTMVTAQGPATLGPGKRLYGLMPNNNLGWVDERM- VDGEMRPRMSAELSRVI G 225 ruler . . . . . . . 160. . . . . . . 170. . . . . . . 180. . . . . . . 190. . . . . . . 200. . . . . . . 210. . . . . . . 220. . . . . . . 230. . . . . . . 240
Rv0813c, a hypothetical protein conservedin mycobacteria and corynebacteria
M. tuberculosis M. leprae
3D Structure Resolution• Strategies
– Absence of methionines• Double mutant Ile->Met
– Anomalous diffraction experiments• SAD & MAD on ID29• Selenium K-edge• Se(Met) crystal size < 40µm
– Phase extension to 1.7Å– Automatic tracing
• 80% amino acids
Rv0813c, a FABP-like fold
Top viewSide view
Ligand Binding Pocket
XY HOH2O
Tyr 192
ML2640 gene family * . : * * * : : : . : . : : . ML2640 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTHDDTWDI KTSVGTTAVMVAAARAAETDRPDALI RDPYAKLLVTNTGAGALWEAMLDPSMVAKVEAI DAEAAAMVEHMRSYQAVRTNFFDTYFNNAVI DGI RQFV 107 Rv0146 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTHDDTWDI KTSVGATAVMVAAARAVETDRPDPLI RDPYARLLVTNAGAGAI WEAMLDPTLVAKAAAI DAETAAI VAYLRSYQAVRTNFFDTYFASAVAAGI RQVV 107 Rv0145 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTELDDVSSLPSSRRTAGDTWAI TESVGATALGVAAARAVETAATNPLI RDEFAKVLVS- - SAGTAWARLADADLAWLDG- - DQLGRRVHRVACDYQAVRTHFFDEYFGAAVDAGVRQVV 116 Rv1896c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTTPEYGSLRSDDDHWDI VSNVGYTALLVAGWRALHTTGPKPLVQDEYAKHFI T- - ASA- - - DPYLEGLLAN- - PRTSEDGTAFPR- - - - LYGVQTRFFDDFFNCADEAGI RQAV 104 ML2020 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MMAATPEFGSLRSDDDHWDI VSSVGYTALLVAGWRALHAVGPQPLVRDEYAKYFI T- - ASR- - - DPYLMNLLAN- - PGTSLNETAFPR- - - - LYGVQTRFFDDFFSSAGDTGI RQAV 106 Rv0893c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTEDDSWDVTTSVGSTGLLVAAARALETQKADPLAI DPYAEVFCR- - AAGGEWADVLDGKLPDHYLTTGDFGEHFVN- - - - FQGARTRYFDEYFSRATAAGMKQVV 101 Rv0281 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTEGDSWDI TTSVGSTALFVATARALEAQKSDPLVVDPYAEAFCR- - AVGGSWADVLDGKLPDHKLKSTDFGEHFVN- - - - FQGARTKYFDEYFRRAAAAGARQVV 101 Rv3767c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MPRTDNDSWAI TESVGATALGVAAARAAETESDNPLI NDPFARI FVD- AAGDGI WSMYTNRTLLAGATDLDPDLRAPI QQMI DFMAARTAFFDEYFLATADAGVRQVV 107 Rv1729c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MARTDDDNWDLTSSVGVTATI VAVGRALATKDPRGLI NDPFAEPLVR- AVGLDLFTKMMDGELDMSTI ADVSPA- - VAQAMVYGNAVRTKYFDDYLLNATAGGI RQVA 105 Rv0725c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MPRAHDDNWDLASSVGATATMVAAGRALATKDPRGLI NDPFAEPLVR- AVGLDFFTKLI DGELDI ATTGNLSPG- - RAQAMI DGI AVRTKYFDDYFRTATDGGVRQVV 105 Rv0830 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MVRADRDRWDLATSVGATATMVAAQRALAADPRYALI DDPYAAPLVR- AVGMDVYTRLVDWQI PVE- - GDSEFD- - - PQRMATGMACRTRFFDQFFLDATHSGI GQFV 102 Rv3399 MARPMGKLPSNTRKCAQCAMAEALLEI AGQTI NQKDLGRSGRMTRTDNDTWDLASSVGATATMI ATARALASRAENPLI NDPFAEPLVR- AVGI DLFTRLASGELRLEDI GDHATG- - - GRWMI DNI AI RTKFYDDFFGDATTAGI RQVV 146 Rv0726c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTYTGSI RCEGDTWDLASSVGATATMVAAARAMATRAANPLI NDQFAEPLVR- AVGVDVLTRLASGELTASDI DDPERPNASMVRMAEHHAVRTKFFDEFFMDATRAGI RQVV 112 Rv0731c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTQTGSARFEGDSWDLASSVGLTATMVAAARAVAGRAPGALVNDQFAEPLVR- AVGVDFFVRMASGELDPDELAEDEAN- - GLRRFADAMAI RTHYFDNFFLDATRAGI RQAV 110 Rv3787c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MARTDDDSWDLATGVGATATLVAAGRARAARAAQPLI DDPFAEPLVR- AVGVEFLTRWATGELDAADVDDPDAA- WGLQRMTTELVVRTRYFDQFFLDAAAAGVRQAV 106 Rv2751 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MARN- - - - - - - PAAQTAFGPMVLAAVEQNEPPGRRLVDDDLADLFLP- RPLRWLAGATRSAVLRRLLI SASEWS- - - GRGLWANLACRKRFI GDKLDEALGD- I DAVV 96 ruler 1. . . . . . . 10. . . . . . . . 20. . . . . . . . 30. . . . . . . . 40. . . . . . . . 50. . . . . . . . 60. . . . . . . . 70. . . . . . . . 80. . . . . . . . 90. . . . . . . 100. . . . . . . 110. . . . . . . 120. . . . . . . 130. . . . . . . 140. . . . . . . 150
*: . : *** *. : ** : : *: * * : * : : *: . : . * * : **: ** . ML2640 I LASGLDSRAYRLDWPTGTTVYEI DQPKVLAYKSTTLAEHGVTPTADRREVPI DLR- QDWPPALRSAGFDPSARTAWLAEGLLMYLPATAQDGLFTEI GGLSAVGSRI AVETSPLHGDE- - - - WREQMQLRFRRVSDALGFEQAVDVQEL 252 Rv0146 I LASGLDSRAYRLDWPAGTI VYEI DQPKVLSYKSTTLAENGVTPSAGRREVPADLR- QDWPAALRDAGFDPTARTAWLAEGLLMYLPAEAQDRLFTQVGAVSVAGSRI AAETAPVHGEE- - - - RRAEMRARFKKVADVLGI EQTI DVQEL 252 Rv0145 I LAAGLDARAYRLNWPAGTVVYEI DQPSVLEYKAGI LQSHGAVPTARRHAVAVDLR- DDWPAALI AAGFDGTQPTAWLAEGLLPYLPGDAADRLFDMVTALSAPGSQVAVEAFTMN- - - - - - - - TKGNTQRWNRMRERLGLD- - I DVQAL 255 Rv1896c I VAAGLDCRAYRLDWQPGTTVFEI DVPKVLEFKARVLSERGAVPKAHRVAVPADLR- TDWPTPLTAAGFDPQRPSAWSVEGLLPYLTGDAQYALFARI DELCAPGSRVALGALGSRLDH- - - - EQLAALETAHPGVNMSG- - - DVNFSAL 246 ML2020 I VAAGLDSRAYRLKWPNGATVFEI DLPKVLEFKARVLAEQGAI PNAGRSEVAADLR- ADWPRALKAAGFDPQRSSAWSVEGLLPYLTNDAQSALFTRI GELCAPGSRI AVGALGSRLDR- - - - KQLAALEATHPGVNI SG- - - DVDFSAL 248 Rv0893c I LAAGLDSRAFRLQWPI GTTI FELDRPQVLDFKNAVLADYHI RPRAQRRSVAVDLR- DEWQI ALCNNGFDANRPSAWI AEGLLVYLSAEAQQRLFI GI DTLASPGSHVAV- EEATPLDP- - - - CEFAAKLERERAANAQGDP- RR- FFQM 243 Rv0281 I LAAGLDSRAYRLPWPDGTTVFELDRPQVLDFKREVLASHGAQPRALRREI AVDLR- DDWPQALRDSGFDAAAPSAWI AEGLLI YLPATAQERLFTGI DALAGRRSHVAV- EDGAPMGP- - - - DEYAAKVEEERAAI AEGAE- EHPFFQL 244 Rv3767c I LASGLDSRAWRLPWPDGTVVYELDQPKVLEFKSATLRQHGAQPASQLVNVPI DLR- QDWPKALQKAGFDPSKPCAWLAEGLVRYLPARAQDLLFERI DALSRPGSWLASNVPGAGFLDPERMRRQRADMRRMRAAAAKLVETEI SDVDD 256 Rv1729c I LASGLDSRAYRLPWPTRTVVYEI DQPKVMEFKTTTLADLGAEPSAI RRAVPI DLR- ADWPTALQAAGFDSAAPTAWLAEGLLI YLKPQTQDRLFDNI TALSAPGSMVATEFVTGI ADFSA- - - E- - - - RARTI SNPFRCHGVDVDLASL 247 Rv0725c I LAAGLDARAYRLPWPAGTVVYEI DQPQVI DFKTTTLAGI GAKPTAI RRTVYI DLR- ADWPAALQAAGLDSTAPTAWLAEGMLI YLPPDPRTG- - - - - CSTTAPNSVLR- - AARSLPNLS- - - - - - - - - RALWI STQAGYEKWRI RFAST 238 Rv0830 I LASGLDARAYRLAWPVGSI VYEVDMPEVI EFKTATLSDLGAEPATERRTVAVDLR- DDWATALQTAGFDPKVPAAWSAEGLLVYLPVEAQDALFDNI TALSAPGSRLAFE- - FVPDTAI F- - - ADERWRN- - YHNRMSELGFDI DLNEL 244 Rv3399 I LAAGLDTRAYRLPWPPGTVVYEI DQPAVI KFKTRALANLNAEPNAERHAVAVDLR- NDWPTALKNAGFDPARPTAFSAEGLLSYLPPQGQDRLLDAI TALSAPDSRLATQSPLVLDLAEE- - - DEKKMRMKSAAEAWRERGFDLDLTEL 292 Rv0726c I LASGLDSRAYRLAWPAQTVVYEI DQPQVMEFKTRTLAELGATPTADRRVVTADLR- ADWPTALGAAGFDPTQPTAWSAEGLLRYLPPEAQDRLLDNVTALSVPDSRFATESI RNFKPHHE- - - ERMRERMTI LANRWRAYGFDLDMNEL 258 Rv0731c I LASGLDSRAYRLRWPAGTI VFEVDQPQVI DFKTTTLAGLGAAPTTDRRTVAVDLR- DDWPTALQKAGFDNAQRTAWI AEGLLGYLSAEAQDRLLDQI TAQSVPGSQFATEVLRDI NRLNE- - - EELRGRMRRLAERFRRHGLDLDMSGL 256 Rv3787c I LASGLDARGYRLPWPADTTVFEVDQPRVLEFKAQTLAGLGAQPTADLRMVPADLR- HDWPDALRRGGFDAAEPAAWI AEGLFGYLPPDAQNRLLDHVTDLSAPGSRLALEAFLGSADRDS- - - ARVEEMI RTATRGWREHGFHLDI WAL 252 Rv2751 I LGAGLDTRAYRLTRRVRMPVFEVDLPVNI ARKAKTVRRVLGELPLSVRLVALDFEHDDLLTALAEHGYRTEYRVFFVCEGVTQYLTERAVRRTLEGLRAAAPGSRMVFTYVRRDFI DG- - - - - - - - - - - - TNRYGTRTLYHTVRQRRQL 234 ruler . . . . . . . 160. . . . . . . 170. . . . . . . 180. . . . . . . 190. . . . . . . 200. . . . . . . 210. . . . . . . 220. . . . . . . 230. . . . . . . 240. . . . . . . 250. . . . . . . 260. . . . . . . 270. . . . . . . 280. . . . . . . 290. . . . . . . 300
ML2640 I YHDENRAVVADWLNRHGWRATAQSAPDEMRRVGRWGDGVPMADDKD- - - - AFAEFVTAHRL- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 310 Rv0146 VYHDQDRASVADWLTDHGWRARSQRAPDEMRRVGRWVEGVPMADDPT- - - - AFAEFVTAERL- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 310 Rv0145 TYHEPDRSDAAQWLATHGWQVHSVSNREEMARLGRAI P- QDLVDETVRTTLLRGRLVTPAQPA- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 317 Rv1896c TYD- DKTD- PVEWLVEHGWAVDPVRSTLELQVGYGLTPPDVDVKI DS- - - FMRSQYI TAVRA- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 303 ML2020 TYE- PKTD- SAQWLAAHGWAVEPVRNTLELQTSYGMTPPDVDVQMDS- - - FMHSQYI TATR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 304 Rv0893c VYN- ERWARATEWFDERGWRATATPLAEYLRRVGRAVPEADTEAAPM- - - VTAI TFVSAVRTGLVADPARTSPSSTSI GFKRFEAD- - - - - - - - - - - - - - - - - - - - - - - - - 325 Rv0281 VYN- ERCAPAAEWFGERGWTAVATLLNDYLEAVGRPVPGPESEAGPM- - - FARNTLVSAARV- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 302 Rv3767c LWYAEQRTAVAEWLRERGWDVSTATLPELLARYGRSI PHSGEDSI PP- - - - - - NLFVSAQRATS- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 314 Rv1729c VYT- GPRNHVLDYLAAKGWQPEGVSLAELFRRSGLDVRAADDDTI FI SGCLTDHSSI SPPTAAGWR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 312 Rv0725c AWT- STWRRWCI PANAATSSTTCAPRAGTLRAQCGPTYSGAMVCPFPPHTTTI RSAKSSSSAVV- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 301 Rv0830 VYH- GQRGHVLDYLTRDGWQTSALTVTQLYEANGFAYPDDELATAFADLTYSSATLMR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 301 Rv3399 I YF- DQRNDVADYLAGSGWQVTTSTGKELFAAQGLPPFADDHI TRFADRRYI SAVLK- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 348 Rv0726c VYF- GDRNEPASYLSDNGWLLTEI KSQDLLTANGFQPFEDEEVPLPD- FFYVSARLQRKHRQYPAHRKPAPSWRHTACPVNELSKSAAYTMTRSDAHQASTTAPPPPGLTG 367 Rv0731c VYF- GDRTDARTYLADHGWRTASASTTDLLAEHGLPPI DGDDAPFGE- VI YVSAELKQKHQDTR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 318 Rv3787c NYA- GPRHEVSGYLDNHGWRSVGTTTAQLLAAHDLPAAPALPAGLADRPNYWTCVLG- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 308 Rv2751 WHFGLDPEEVAGFLADYGWRLTEQAGPEELVQRYVEPTGRNLNASQI EWSAYAEKSEPVTPR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 296 ruler . . . . . . . 310. . . . . . . 320. . . . . . . 330. . . . . . . 340. . . . . . . 350. . . . . . . 360. . . . . . . 370. . . . . . . 380. . . . . . . 390. . . . . . . 400. . . . . . . 410.
~50%2.8 Å1531H1D
~50%3.1 Å1731IM8
~70%2.2 Å2181RJE
SSEr.m.s.deviation
Equiv.residues
PDB
1RJE 1IM8 1H1DLeu C-methyltransferase (PPM1) YecO H. influenzae (AdoMet) Catechol O-methyltransferase (rat)
ML2640
ML2640
In progress: search for TB substrate(s)
* . : * * * : : : . : . : : . ML2640 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTHDDTWDI KTSVGTTAVMVAAARAAETDRPDALI RDPYAKLLVTNTGAGALWEAMLDPSMVAKVEAI DAEAAAMVEHMRSYQAVRTNFFDTYFNNAVI DGI RQFV 107 Rv0146 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTHDDTWDI KTSVGATAVMVAAARAVETDRPDPLI RDPYARLLVTNAGAGAI WEAMLDPTLVAKAAAI DAETAAI VAYLRSYQAVRTNFFDTYFASAVAAGI RQVV 107 Rv0145 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTELDDVSSLPSSRRTAGDTWAI TESVGATALGVAAARAVETAATNPLI RDEFAKVLVS- - SAGTAWARLADADLAWLDG- - DQLGRRVHRVACDYQAVRTHFFDEYFGAAVDAGVRQVV 116 Rv1896c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTTPEYGSLRSDDDHWDI VSNVGYTALLVAGWRALHTTGPKPLVQDEYAKHFI T- - ASA- - - DPYLEGLLAN- - PRTSEDGTAFPR- - - - LYGVQTRFFDDFFNCADEAGI RQAV 104 ML2020 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MMAATPEFGSLRSDDDHWDI VSSVGYTALLVAGWRALHAVGPQPLVRDEYAKYFI T- - ASR- - - DPYLMNLLAN- - PGTSLNETAFPR- - - - LYGVQTRFFDDFFSSAGDTGI RQAV 106 Rv0893c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTEDDSWDVTTSVGSTGLLVAAARALETQKADPLAI DPYAEVFCR- - AAGGEWADVLDGKLPDHYLTTGDFGEHFVN- - - - FQGARTRYFDEYFSRATAAGMKQVV 101 Rv0281 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MRTEGDSWDI TTSVGSTALFVATARALEAQKSDPLVVDPYAEAFCR- - AVGGSWADVLDGKLPDHKLKSTDFGEHFVN- - - - FQGARTKYFDEYFRRAAAAGARQVV 101 Rv3767c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MPRTDNDSWAI TESVGATALGVAAARAAETESDNPLI NDPFARI FVD- AAGDGI WSMYTNRTLLAGATDLDPDLRAPI QQMI DFMAARTAFFDEYFLATADAGVRQVV 107 Rv1729c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MARTDDDNWDLTSSVGVTATI VAVGRALATKDPRGLI NDPFAEPLVR- AVGLDLFTKMMDGELDMSTI ADVSPA- - VAQAMVYGNAVRTKYFDDYLLNATAGGI RQVA 105 Rv0725c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MPRAHDDNWDLASSVGATATMVAAGRALATKDPRGLI NDPFAEPLVR- AVGLDFFTKLI DGELDI ATTGNLSPG- - RAQAMI DGI AVRTKYFDDYFRTATDGGVRQVV 105 Rv0830 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MVRADRDRWDLATSVGATATMVAAQRALAADPRYALI DDPYAAPLVR- AVGMDVYTRLVDWQI PVE- - GDSEFD- - - PQRMATGMACRTRFFDQFFLDATHSGI GQFV 102 Rv3399 MARPMGKLPSNTRKCAQCAMAEALLEI AGQTI NQKDLGRSGRMTRTDNDTWDLASSVGATATMI ATARALASRAENPLI NDPFAEPLVR- AVGI DLFTRLASGELRLEDI GDHATG- - - GRWMI DNI AI RTKFYDDFFGDATTAGI RQVV 146 Rv0726c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTYTGSI RCEGDTWDLASSVGATATMVAAARAMATRAANPLI NDQFAEPLVR- AVGVDVLTRLASGELTASDI DDPERPNASMVRMAEHHAVRTKFFDEFFMDATRAGI RQVV 112 Rv0731c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MTQTGSARFEGDSWDLASSVGLTATMVAAARAVAGRAPGALVNDQFAEPLVR- AVGVDFFVRMASGELDPDELAEDEAN- - GLRRFADAMAI RTHYFDNFFLDATRAGI RQAV 110 Rv3787c - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MARTDDDSWDLATGVGATATLVAAGRARAARAAQPLI DDPFAEPLVR- AVGVEFLTRWATGELDAADVDDPDAA- WGLQRMTTELVVRTRYFDQFFLDAAAAGVRQAV 106 Rv2751 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - MARN- - - - - - - PAAQTAFGPMVLAAVEQNEPPGRRLVDDDLADLFLP- RPLRWLAGATRSAVLRRLLI SASEWS- - - GRGLWANLACRKRFI GDKLDEALGD- I DAVV 96 ruler 1. . . . . . . 10. . . . . . . . 20. . . . . . . . 30. . . . . . . . 40. . . . . . . . 50. . . . . . . . 60. . . . . . . . 70. . . . . . . . 80. . . . . . . . 90. . . . . . . 100. . . . . . . 110. . . . . . . 120. . . . . . . 130. . . . . . . 140. . . . . . . 150
*: . : *** *. : ** : : *: * * : * : : *: . : . * * : **: ** . ML2640 I LASGLDSRAYRLDWPTGTTVYEI DQPKVLAYKSTTLAEHGVTPTADRREVPI DLR- QDWPPALRSAGFDPSARTAWLAEGLLMYLPATAQDGLFTEI GGLSAVGSRI AVETSPLHGDE- - - - WREQMQLRFRRVSDALGFEQAVDVQEL 252 Rv0146 I LASGLDSRAYRLDWPAGTI VYEI DQPKVLSYKSTTLAENGVTPSAGRREVPADLR- QDWPAALRDAGFDPTARTAWLAEGLLMYLPAEAQDRLFTQVGAVSVAGSRI AAETAPVHGEE- - - - RRAEMRARFKKVADVLGI EQTI DVQEL 252 Rv0145 I LAAGLDARAYRLNWPAGTVVYEI DQPSVLEYKAGI LQSHGAVPTARRHAVAVDLR- DDWPAALI AAGFDGTQPTAWLAEGLLPYLPGDAADRLFDMVTALSAPGSQVAVEAFTMN- - - - - - - - TKGNTQRWNRMRERLGLD- - I DVQAL 255 Rv1896c I VAAGLDCRAYRLDWQPGTTVFEI DVPKVLEFKARVLSERGAVPKAHRVAVPADLR- TDWPTPLTAAGFDPQRPSAWSVEGLLPYLTGDAQYALFARI DELCAPGSRVALGALGSRLDH- - - - EQLAALETAHPGVNMSG- - - DVNFSAL 246 ML2020 I VAAGLDSRAYRLKWPNGATVFEI DLPKVLEFKARVLAEQGAI PNAGRSEVAADLR- ADWPRALKAAGFDPQRSSAWSVEGLLPYLTNDAQSALFTRI GELCAPGSRI AVGALGSRLDR- - - - KQLAALEATHPGVNI SG- - - DVDFSAL 248 Rv0893c I LAAGLDSRAFRLQWPI GTTI FELDRPQVLDFKNAVLADYHI RPRAQRRSVAVDLR- DEWQI ALCNNGFDANRPSAWI AEGLLVYLSAEAQQRLFI GI DTLASPGSHVAV- EEATPLDP- - - - CEFAAKLERERAANAQGDP- RR- FFQM 243 Rv0281 I LAAGLDSRAYRLPWPDGTTVFELDRPQVLDFKREVLASHGAQPRALRREI AVDLR- DDWPQALRDSGFDAAAPSAWI AEGLLI YLPATAQERLFTGI DALAGRRSHVAV- EDGAPMGP- - - - DEYAAKVEEERAAI AEGAE- EHPFFQL 244 Rv3767c I LASGLDSRAWRLPWPDGTVVYELDQPKVLEFKSATLRQHGAQPASQLVNVPI DLR- QDWPKALQKAGFDPSKPCAWLAEGLVRYLPARAQDLLFERI DALSRPGSWLASNVPGAGFLDPERMRRQRADMRRMRAAAAKLVETEI SDVDD 256 Rv1729c I LASGLDSRAYRLPWPTRTVVYEI DQPKVMEFKTTTLADLGAEPSAI RRAVPI DLR- ADWPTALQAAGFDSAAPTAWLAEGLLI YLKPQTQDRLFDNI TALSAPGSMVATEFVTGI ADFSA- - - E- - - - RARTI SNPFRCHGVDVDLASL 247 Rv0725c I LAAGLDARAYRLPWPAGTVVYEI DQPQVI DFKTTTLAGI GAKPTAI RRTVYI DLR- ADWPAALQAAGLDSTAPTAWLAEGMLI YLPPDPRTG- - - - - CSTTAPNSVLR- - AARSLPNLS- - - - - - - - - RALWI STQAGYEKWRI RFAST 238 Rv0830 I LASGLDARAYRLAWPVGSI VYEVDMPEVI EFKTATLSDLGAEPATERRTVAVDLR- DDWATALQTAGFDPKVPAAWSAEGLLVYLPVEAQDALFDNI TALSAPGSRLAFE- - FVPDTAI F- - - ADERWRN- - YHNRMSELGFDI DLNEL 244 Rv3399 I LAAGLDTRAYRLPWPPGTVVYEI DQPAVI KFKTRALANLNAEPNAERHAVAVDLR- NDWPTALKNAGFDPARPTAFSAEGLLSYLPPQGQDRLLDAI TALSAPDSRLATQSPLVLDLAEE- - - DEKKMRMKSAAEAWRERGFDLDLTEL 292 Rv0726c I LASGLDSRAYRLAWPAQTVVYEI DQPQVMEFKTRTLAELGATPTADRRVVTADLR- ADWPTALGAAGFDPTQPTAWSAEGLLRYLPPEAQDRLLDNVTALSVPDSRFATESI RNFKPHHE- - - ERMRERMTI LANRWRAYGFDLDMNEL 258 Rv0731c I LASGLDSRAYRLRWPAGTI VFEVDQPQVI DFKTTTLAGLGAAPTTDRRTVAVDLR- DDWPTALQKAGFDNAQRTAWI AEGLLGYLSAEAQDRLLDQI TAQSVPGSQFATEVLRDI NRLNE- - - EELRGRMRRLAERFRRHGLDLDMSGL 256 Rv3787c I LASGLDARGYRLPWPADTTVFEVDQPRVLEFKAQTLAGLGAQPTADLRMVPADLR- HDWPDALRRGGFDAAEPAAWI AEGLFGYLPPDAQNRLLDHVTDLSAPGSRLALEAFLGSADRDS- - - ARVEEMI RTATRGWREHGFHLDI WAL 252 Rv2751 I LGAGLDTRAYRLTRRVRMPVFEVDLPVNI ARKAKTVRRVLGELPLSVRLVALDFEHDDLLTALAEHGYRTEYRVFFVCEGVTQYLTERAVRRTLEGLRAAAPGSRMVFTYVRRDFI DG- - - - - - - - - - - - TNRYGTRTLYHTVRQRRQL 234 ruler . . . . . . . 160. . . . . . . 170. . . . . . . 180. . . . . . . 190. . . . . . . 200. . . . . . . 210. . . . . . . 220. . . . . . . 230. . . . . . . 240. . . . . . . 250. . . . . . . 260. . . . . . . 270. . . . . . . 280. . . . . . . 290. . . . . . . 300
ML2640 I YHDENRAVVADWLNRHGWRATAQSAPDEMRRVGRWGDGVPMADDKD- - - - AFAEFVTAHRL- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 310 Rv0146 VYHDQDRASVADWLTDHGWRARSQRAPDEMRRVGRWVEGVPMADDPT- - - - AFAEFVTAERL- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 310 Rv0145 TYHEPDRSDAAQWLATHGWQVHSVSNREEMARLGRAI P- QDLVDETVRTTLLRGRLVTPAQPA- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 317 Rv1896c TYD- DKTD- PVEWLVEHGWAVDPVRSTLELQVGYGLTPPDVDVKI DS- - - FMRSQYI TAVRA- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 303 ML2020 TYE- PKTD- SAQWLAAHGWAVEPVRNTLELQTSYGMTPPDVDVQMDS- - - FMHSQYI TATR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 304 Rv0893c VYN- ERWARATEWFDERGWRATATPLAEYLRRVGRAVPEADTEAAPM- - - VTAI TFVSAVRTGLVADPARTSPSSTSI GFKRFEAD- - - - - - - - - - - - - - - - - - - - - - - - - 325 Rv0281 VYN- ERCAPAAEWFGERGWTAVATLLNDYLEAVGRPVPGPESEAGPM- - - FARNTLVSAARV- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 302 Rv3767c LWYAEQRTAVAEWLRERGWDVSTATLPELLARYGRSI PHSGEDSI PP- - - - - - NLFVSAQRATS- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 314 Rv1729c VYT- GPRNHVLDYLAAKGWQPEGVSLAELFRRSGLDVRAADDDTI FI SGCLTDHSSI SPPTAAGWR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 312 Rv0725c AWT- STWRRWCI PANAATSSTTCAPRAGTLRAQCGPTYSGAMVCPFPPHTTTI RSAKSSSSAVV- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 301 Rv0830 VYH- GQRGHVLDYLTRDGWQTSALTVTQLYEANGFAYPDDELATAFADLTYSSATLMR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 301 Rv3399 I YF- DQRNDVADYLAGSGWQVTTSTGKELFAAQGLPPFADDHI TRFADRRYI SAVLK- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 348 Rv0726c VYF- GDRNEPASYLSDNGWLLTEI KSQDLLTANGFQPFEDEEVPLPD- FFYVSARLQRKHRQYPAHRKPAPSWRHTACPVNELSKSAAYTMTRSDAHQASTTAPPPPGLTG 367 Rv0731c VYF- GDRTDARTYLADHGWRTASASTTDLLAEHGLPPI DGDDAPFGE- VI YVSAELKQKHQDTR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 318 Rv3787c NYA- GPRHEVSGYLDNHGWRSVGTTTAQLLAAHDLPAAPALPAGLADRPNYWTCVLG- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 308 Rv2751 WHFGLDPEEVAGFLADYGWRLTEQAGPEELVQRYVEPTGRNLNASQI EWSAYAEKSEPVTPR- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 296 ruler . . . . . . . 310. . . . . . . 320. . . . . . . 330. . . . . . . 340. . . . . . . 350. . . . . . . 360. . . . . . . 370. . . . . . . 380. . . . . . . 390. . . . . . . 400. . . . . . . 410.
1RJELeulliot et al, J.Biol.Chem., 2004
GPH - Tuberculose (IP)
Structural Genomics
Opportunistic approachBiological relevance of
structuresProtein complexes
Methodology bottlenecks(membrane proteins)
New unique foldsReduced costsTechnology
developmentsFunctional annotation
Drug discovery
AcknowledgementsThe pipelineJacques BellalouVincent BondetCedric Fiez-VandalFabrice GuillemotAhmed HaouzNadine HonoréStéphane PetresFlorence Proux
Bill Shepard (ESRF)
Research labsStewart Cole (UGMB, IP) Mycobacterial genomics
Brigitte Gicquel (UGM, IP) Gene essentiality
Jean M. Betton (URMP, IP) Protein expression
Muriel Delepierre (URMN, IP) NMR
Michael Nilges (UBS, IP) Structural bioinformatics
Pedro M. Alzari (UBS, IP) Protein crystallography
Funding: Inst. Pasteur, MENRT, French Genopole, X-TB, SPINE