200
DESCRIPTION
b 4 451.2. - 164 805.4. (A). - 164 - 98 707.4. 100. y 1 222.2. (B). 476.5. 100. 632.4. b 7 748.3. PP + H + 165.1. 689.5. 787.5. 730.3. b 7 - 98 650.4. Relative Abundance. 50. (y 6 – 98) 2+ 337.5. Relative Abundance. 50. 787.4. PP + H + 165.1. y 4 - PowerPoint PPT PresentationTRANSCRIPT
Supplemental Information SF1. CAD spectra of doubly charged peptide TPpTAPSLG after derivatization with the following reagents: (A) and (B) 1-(2-pyrimidyl)piperazine (PP) with RT 13.2 min (A) and 14.6 min (B), respectively, (C) 2-nitrophenylhydrazine (2-NPH), (D) 3-nitrophenylhydrazine hydrochloride (3-NPH.HCl), and (E) 4-nitrophenylhydrazine (4-NPH).
200 400 600 800m/z
0
50
100
Rel
ativ
e A
bund
ance
- 164805.4
- 164 - 98707.5
787.5
PP + H+
165.1
(A)
200 400 600 800m/z
0
50
100
Rel
ativ
e A
bund
ance
(B) y1
222.2
- 164805.3
787.4
PP + H+
165.1
- 164 - 98707.4
b7
748.3
730.3
689.5b7 - 98
650.4
632.4
y4
519.4
476.5
b4
451.2
(y6 – 98)2+
337.5
200 400 600 800m/z
0
50
100
Rel
ativ
e A
bund
ance
(C)
y1
211.0
470.9
( – 153)2+
403.4
b7
748.2
( – 98)2+
430.9
y2
324.3
b4
451.1
y4
508.2
b5
548.1
b6
635.3
b7 - 98650.4
b7
748.3
NPH + H+
152.0
200 400 600 800m/z
0
50
100
Rel
ativ
e A
bund
ance
(D)
y1
211.0194.1y2
324.3
( – 153)2+
403.4
( – 98)2+
431.0
b6
635.3
b7 - 98650.4
b5
548.1
470.9
b4
451.2
y4
508.2
200 400 600 800m/z
0
50
100
Rel
ativ
e A
bund
ance
(E)
b7
748.3y1
211.0
194.1
y2
324.3
( – 153)2+
403.4
( – 98)2+
431.0
b6
635.3
b7 - 98650.4
b5
548.1
470.9
b4
451.2
y4
508.3
Supplemental Information SF2. CAD spectra of singly (A) and doubly charged (B) peptide TPTAPpSLG after derivatization with 2-nitrophenylhydrazine (2-NPH).
300 500 700 900 1100m/z
0
50
100
Rel
ativ
e A
bund
ance
940.3
– 98860.5
b7
748.4
y4
588.5
y6
760.4
y5
659.4
b7 - 98650.5
730.4
632.5
y4 - 98490.5
(A)
y1
211.1
( – 98)2+
430.9
b4
371.2
200 400 600 800 1000m/z
0
50
100
Rel
ativ
e A
bund
ance
(B)
y4
588.2
b7
748.2
b5
468.3
Supplemental Information SF3. CAD spectra of singly (A) and doubly charged (B) peptide SIINFEKL after derivatization with 2-nitrophenylhydrazine (2-NPH).
300 500 700 900 1100m/z
0
50
100
Rel
ativ
e A
bund
ance
1081.5
– 153945.6
b7
832.5
y2
395.5
y6
898.5
b5
575.5
927.7
557.5
(A)
y3
524.5
y5
785.6
687.5
b6
704.5
y4
671.5
b7
832.4
y5
785.5
200 400 600 800 1000 1200m/z
0
50
100
Rel
ativ
e A
bund
ance
(B)y6
898.4
815.4
b6
704.4
687.4
y4
671.5
b5
575.4
558.3
y3
524.4
541.3
(-153)2+
473.6
(-153 - 28)2+
459.7
b3
314.3
y2
395.4
b72+
417.1
y1
267.2
a2
173.2
b2
201.2
250.3
707.5
b8
899.5
1128.5
y6
725.5y8
910.5b9
1000.5
b9 + 181018.4
b7
784.5
1111.5
(A)
400 600 800 1000 1200 1400m/z
50
100R
elat
ive
Abu
ndan
ce
0400 600 800 1000 1200 1400
m/z
(B)
50
100
Rel
ativ
e A
bund
ance
0
b8
899.5
y6
860.5
b7
784.5
- 1531128.5
y8
1045.5
1263.5
b9
1000.4
400 600 800 1000 1200 1400m/z
(C)
50
100
0
1199.6
- 1531263.6
y8
1180.5
b8
1034.4
y6
995.5b7
784.5
b9
1135.5
Rel
ativ
e A
bund
ance
Supplemental Information SF4. CAD spectra of singly charged peptide VHGQPHQDTK: (A) without derivatization, (B) and (C) after derivatization with 2-nitrophenylhydrazine (2-NPH). (B) mono-NPHylation, (C) di-NPHylation.
Supplemental Information SF5. CAD spectra of doubly charged peptide EHAGVISVL after derivatization with (A) 3-nitrophenylhydrazine hydrochloride (3-NPH.HCl) and (B) 4-nitrophenylhydrazine (2-NPH).
200 400 600 800 1000m/z
0
50
100
Rel
ativ
e A
bund
ance
(-153 - 28)2+
440.5
b8
793.5
757.5
b6
607.5
775.5
y4
566.5
676.4
b5
494.4389.2
(-153)2+
454.2
b7
694.5
y2
366.8
b82+
398.0
y1
267.5
250.6
b08 - I
662.4
589.5
521.4
(A)
200 400 600 800 1000m/z
0
50
100
Rel
ativ
e A
bund
ance
b8
793.4
757.4
b6
607.4
775.4
y4
566.3
676.7
b5
495.4
388.7
(-153)2+
454.1
b7
694.4
y2
366.4
b82+
397.5y1
267.2
250.3
b08 - I
662.3
589.3
521.4
(B)
Supplemental Information SF6. CAD spectra of singly charged peptide EHAGVISVL: (A) without derivatization, and after derivatization with (B) 2-nitrophenylhydrazine (2-NPH), (C) 3-nitrophenylhydrazine hydrochloride (3-NPH.HCl), and (D) 4-nitrophenylhydrazine (4-NPH).
300 500 700 900 1100m/z
0
50
100
Rel
ativ
e A
bund
ance
(B)
b5
494.4- 153906.5
775.5b6
607.5
b8
793.5
1041.5
888.5
476.3
589.4
757.4
300 500 700 900 1100m/z
0
50
100
Rel
ativ
e A
bund
ance
(D)
b5
494.4
- 153906.6
775.6b6
607.5
b8
793.5
1041.5
888.6
589.4
757.5
476.4
300 500 700 900 1100m/z
0
50
100
Rel
ativ
e A
bund
ance
b5
494.5
b7
694.7
775.7
b6
607.7
b8
793.6
906.6
(A)
757.6
888.7
476.6
589.6
676.7
y7
658.7
300 500 700 900 1100m/z
0
50
100
Rel
ativ
e A
bund
ance
(C)
b5
494.4
- 153906.6
775.6b6
607.5b8
793.6
1041.5
888.6
589.5
757.5
476.4