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Supplementary Figures Accompanying:
Improvement of Capture Compound Mass Spectrometry (CCMS) Technology for the Profiling of Human Kinases by Combination with 2D LC-MS/MS
Jenny J. Fischer, Olivia Graebner, Mathias Dreger, Mirko Glinski, Sabine Baumgart, and Hubert Koester
Analytics department, caprotec bioanalytics GmbH, Volmerstrasse 5, 12489 Berlin, Germany
1
MS spectra of kinases identified by one single peptide
PRP4B_HUMAN
401,3?479,3?
498,5?
507,6?
y10-NH3-H2O+2Hb10-NH3-H2O+2Hy8+1
parent+2H-H2O
parent+2H-NH3
b2y1
b3y2 b4 y3
y4
b5y5
b6b7
y6b8 y7 b9
y8A V G D E M G T V R
R V T G M E D G V A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.033,48 AMU, +2 H (Parent Error: -13 ppm)
AAPK2_HUMAN
327,2?
762,3?974,5?y10-H2O-H2O+2H
b10-H2O-H2O+2H
b10-H2O+2H
a8+1
parent+2H-H2O
parent+2H-NH3
y1 b2 y2b3 b4
y3
y4
b5y5 b6 b7 y7
b8
y8
b9
S I D D E V V E Q RR Q E V V E D D I S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.188,56 AMU, +2 H (Parent Error: -11 ppm)
NIM1_HUMAN
327,2?
762,3?974,5?y10-H2O-H2O+2H
b10-H2O-H2O+2H
b10-H2O+2H
a8+1
parent+2H-H2O
parent+2H-NH3
y1 b2 y2b3 b4
y3
y4
b5y5 b6 b7 y7
b8
y8
b9
S I D D E V V E Q RR Q E V V E D D I S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.188,56 AMU, +2 H (Parent Error: -11 ppm)
2
Serine/threonine-protein kinase PRP4 homolog OS=Homo sapiens GN=PRPF4B PE=1 SV=2
553,1?
a9-H2O+2H
y9-H2O-H2O+2Hy9-NH3-H2O+2H
parent+2H-H2O
parent+2H-NH3
b2 y2 y3 b4
y4
b5 y5 b6 y6 b7
y7
b8
y8
M T D E E I M E KK E M I E E D T M
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.124,47 AMU, +2 H (Parent Error: -11 ppm)
Tyrosine-protein kinase Fyn OS=Homo sapiens GN=FYN PE=1 SV=3
553,1?
a9-H2O+2H
y9-H2O-H2O+2Hy9-NH3-H2O+2H
parent+2H-H2O
parent+2H-NH3
b2 y2 y3 b4
y4
b5 y5 b6 y6 b7
y7
b8
y8
M T D E E I M E KK E M I E E D T M
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.124,47 AMU, +2 H (Parent Error: -11 ppm)
Hexokinase-1 OS=Homo sapiens GN=HK1 PE=1 SV=3
798,8?
846,4?
854,3?
y15+2H
b9+1
a18-H2O-H2O+2H
y8-H2O
y18-H2O-H2O+2H
b18-NH3-H2O+2Hy11+1
parent+2H-H2O
y3y4 b6
y6
b7
b8
y7
b9
y8
b10
y9
b11
y10
y11
b12
b13
b14y13 b15y14
A V L G P L V G A V D Q G T S S T RR T S S T G Q D V A G V L P G L V A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.726,92 AMU, +2 H (Parent Error: -8,5 ppm)
3
Q59FU6_HUMAN
544,1?
545,4?
553,1?
b5-64b10-H2O-64+2Hy9+2H
b6+0
parent+2H-H2O
parent+2H-NH3
y1b3
y2
b4
y3
b5y4
b6
y5b7
y6
b8
y7b9
y8b10
y9
G A A M+16 V T A V A Y RR Y A V A T V M+16 A A G
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.124,56 AMU, +2 H (Parent Error: -10 ppm)
Misshapen-like kinase 1 OS=Homo sapiens GN=MINK1 PE=1 SV=2
770,4?
787,4?788,4?
b6-NH3+1
y13-NH3-NH3+2H
y2 b3
y3b4
y4b5
y5b6
y6
b7
y7b8
y8 b9 y9 b10y10
b11b12
S L Q Q Q Q Q Q Q Q L Q KK Q L Q Q Q Q Q Q Q Q L S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.611,83 AMU, +2 H (Parent Error: -10,0 ppm)
AURKB_HUMAN
770,4?
787,4?788,4?
b6-NH3+1
y13-NH3-NH3+2H
y2 b3
y3b4
y4b5
y5b6
y6
b7
y7b8
y8 b9 y9 b10y10
b11b12
S L Q Q Q Q Q Q Q Q L Q KK Q L Q Q Q Q Q Q Q Q L S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.611,83 AMU, +2 H (Parent Error: -10,0 ppm)
Creatine kinase B-type OS=Homo sapiens GN=CKB PE=1 SV=1
4
parent+2H-H2O
parent+2H-NH3
b2 y2b3y3 y4 b5 y6 b7 b8
y7
y8
b9y9
b10 y10
V L T P E L Y A E L RR L E A Y L E P T L V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.302,72 AMU, +2 H (Parent Error: -8,4 ppm)
CLK1_HUMAN
y1
b2
y2 b3y3y4 b4
b5
y5
b6
y6
b7y7
I L D F G L A RR A L G F D L I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
903,51 AMU, +2 H (Parent Error: -13 ppm)
B3KU53_HUMAN
b8+1
y15-NH3-H2O+2H
b15-NH3-H2O+2Hy9+1
parent+2H-H2O
y2 b3y3b4
b5y4
b6
y5b7 b8 y7
y8
y9
y10
y11b12y12 b13
A E D G A T P S P S N E T P KK P T E N S P S P T A G D E A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.499,67 AMU, +2 H (Parent Error: -10 ppm)
5
KKCC2_HUMAN
y9+2H
y12-H2O+2H
y5+0
a8
parent+2H-H2O
y1 b4y2
y3
b6
y4
b7
y5
b8
y6
b9
y7 b10y8
b11
y9
y10b12y11 y12
I G G G P G D A A D V Q RR Q V D A A D G P G G G I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.211,58 AMU, +2 H (Parent Error: -12 ppm)
ABL2_HUMAN
a13-NH3-H2O+2Hy13-NH3-NH3+2H
parent+2H-H2O
parent+2H-NH3
y2 b4 y3 b5
y4
b6
b7b8
y7b9
y8
y9
b11 b12
V G E A P G L Q Q P Q P RR P Q P Q Q L G P A E G V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250
1.375,72 AMU, +2 H (Parent Error: -10,0 ppm)
CDK14_HUMAN
a13-NH3-H2O+2Hy13-NH3-NH3+2H
parent+2H-H2O
parent+2H-NH3
y2 b4 y3 b5
y4
b6
b7b8
y7b9
y8
y9
b11 b12
V G E A P G L Q Q P Q P RR P Q P Q Q L G P A E G V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250
1.375,72 AMU, +2 H (Parent Error: -10,0 ppm)
6
HCK_HUMAN
y26-H2O+2H
y26+2H
y26+2H+1
y32-H2O-H2O+2H
y5 y6b5y7 b6y8
y9 y10
b8y11b10y12y13
b12
b13b14y15
b15
y16
y17b17y18y19y20y21b20y22b21y23
y24b22y25b23b24
y26
y27
T S S F A E P G G G G G G G G G G P G G S A S G P G G T G G G KK G G G T G G P G S A S G G P G G G G G G G G G G P E A F S S T
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750 2000 2250
2.432,05 AMU, +2 H (Parent Error: -5,8 ppm)
Tyrosine-protein kinase FRK OS=Homo sapiens GN=FRK PE=1 SV=1
parent+2H-H2Oparent+2H-NH3
y1b2
y2b3
y3
b4
y4
b5
y5
b6
D A W E I P RR P I E W A D
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
885,43 AMU, +2 H (Parent Error: -14 ppm)
Inhibitor of nuclear factor kappa-B kinase subunit epsilon OS=Homo sapiens GN=IKBKE PE=1 SV=1
parent+2H-NH3
b2
y1y2
b3y3 b4 b5
y5
b6 b7
y6
y7
V A D F G L A RR A L G F D A V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
847,45 AMU, +2 H (Parent Error: -15 ppm)
7
M3K2_HUMAN
y6+1
parent+2H-H2O
parent+2H-NH3
y1
b2
y2 y3b3
y4
b4
y5
b5 b6
y6
b7y7
L T D F G A A RR A A G F D T L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
849,43 AMU, +2 H (Parent Error: -15 ppm)
MTOR_HUMAN
parent+2H-H2O
y1 b2 y2 b3 y3b4 y4
y5
y6 b7y7
b8 y8
I A T Q P T N P KK P N T P Q T A I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
968,52 AMU, +2 H (Parent Error: -14 ppm)
Tyrosine-protein kinase JAK3 OS=Homo sapiens GN=JAK3 PE=1 SV=2
b7-H2O-64
a16-64+2H
y15-H2O-H2O+2H
y16+2H
b10-NH3-64a20+2H+1
a11-64b21-H2O-64+2H
a12-64
b3y4
y5
b6
y6
b7
b8
y8 y9b10 y10 b12y12
R S G Q G D A L A S GM+16 P G T E V P G S A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200
2.245,12 AMU, +3 H (Parent Error: -5,7 ppm)
8
AKAP9_HUMAN
b12-NH3-H2O+2H a14+2Ha7-NH3+1
y14-H2O+2H
y14-NH3+2H+1
b7+1
b15-H2O+2H+1
b15-H2O+2H
y7-NH3-NH3a8-NH3-NH3
b15+2H
y16+2H
y16+2H+1a17+2Hy18+2H
b5 y6
b7
y7 y8b9 b10b12
T E Q E A N T E G Q KL E H M K K Q G E T
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200 1400
2.557,23 AMU, +3 H (Parent Error: -8,6 ppm)
Q6LD14_HUMAN
b12-NH3-H2O+2H a14+2Ha7-NH3+1
y14-H2O+2H
y14-NH3+2H+1
b7+1
b15-H2O+2H+1
b15-H2O+2H
y7-NH3-NH3a8-NH3-NH3
b15+2H
y16+2H
y16+2H+1a17+2Hy18+2H
b5 y6
b7
y7 y8b9 b10b12
Q T E Q E A N T E G Q KQ L E H M K K Q G E T N
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
250 500 750 1000 1250 1500
2.557,23 AMU, +3 H (Parent Error: -8,6 ppm)
Nucleoside diphosphate kinase A OS=Homo sapiens GN=NME1 PE=1 SV=1
y7+1y15+2H y8+1y8+2
parent+2H-H2O
y2 b4 y4 b5
y5
b6y6 b7
y7
b8
y8
b9y9
b10
y10
b11
y11b12y12
y14 b16
V M L G E T N P A D S K P G T I RR I T G P K S D A P N T E G L M V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750
1.784,91 AMU, +2 H (Parent Error: -6,8 ppm)
9
ATM_HUMAN
y7+1y15+2H y8+1y8+2
parent+2H-H2O
y2 b4 y4 b5
y5
b6y6 b7
y7
b8
y8
b9y9
b10
y10
b11
y11b12y12
y14 b16
V M L G E T N P A D S K P G T I RR I T G P K S D A P N T E G L M V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750
1.784,91 AMU, +2 H (Parent Error: -6,8 ppm)
Fructosamine-3-kinase OS=Homo sapiens GN=FN3K PE=1 SV=1
y12-H2O-H2O+2Hparent+2H-H2O
b2y2 b3y3
b4 y4 b5y5 y7
y8 b9 y9 b10y10
b11
A Y D T D A G P V F V KK V F V P G A D T D Y A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.281,62 AMU, +2 H (Parent Error: -7,4 ppm)
B4DIG8_HUMAN
y12-H2O-H2O+2Hparent+2H-H2O
b2y2 b3y3
b4 y4 b5y5 y7
y8 b9 y9 b10y10
b11
A Y D T D A G P V F V KK V F V P G A D T D Y A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.281,62 AMU, +2 H (Parent Error: -7,4 ppm)
10
PIM2_HUMAN
a7+1
y24+2H
y24+2H+1
y7 y8 b7 y9b8
b9y10 b10 b11 y12 y16
Q S S T A A A Q S A ST A T D T A S A S Q A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
800 1000 1200 1400
2.701,29 AMU, +3 H (Parent Error: 0,074 ppm)
KAP2_HUMAN
y11+2H
a11-H2O
b9 y8
y9
y10 b12
y11
y12
b19
G P S R C F F G Q V V A A I QG R S H C H Q I A A V V Q G F
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
500 750 1000 1250 1500 1750 2000
3.200,58 AMU, +3 H (Parent Error: -7,0 ppm)
Mitogen-activated protein kinase kinase kinase kinase 5 OS=Homo sapiens GN=MAP4K5 PE=1 SV=1
a7-NH3a16-H2O+2Ha16-NH3+2H
a16-NH3+2H+1
y16-NH3-NH3+2Ha16+2H
y6
b7
y7
b8 y8 b9y9
b10 y10b11 b12
C I V R N P Y T GP N R V I C C K
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
600 800 1000 1200 1400
1.814,85 AMU, +2 H (Parent Error: -6,4 ppm)
11
STK6_HUMAN
a7-NH3a16-H2O+2Ha16-NH3+2H
a16-NH3+2H+1
y16-NH3-NH3+2Ha16+2H
y6
b7
y7
b8 y8b9y9
b10 y10b11 b12
G C H K C C I V R N P Y T G H KK H G T Y P N R V I C C K H C G
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750
1.814,85 AMU, +2 H (Parent Error: -6,4 ppm)
JAK1_HUMAN
y12+2H
b7-NH3-H2Ob7-NH3-NH3
a7a7+1
a14-H2O-H2O+2H
y13+2H
y7+1
y18+2H
y4 y5b5
b6
y6b7
b8 y8
b9 y9 b12
Q P L P S A P E N N PA L E E E P N N E P
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200
2.164,06 AMU, +3 H (Parent Error: -6,7 ppm)
BLK_HUMAN
a11-H2O+2H
a6-64
y11-NH3-H2O+2H
b11-NH3-NH3+2H
b6-NH3-64parent+2H-NH3-64
parent+2H+3-64parent+2H-H2O
y2 b3 b4y4 b5y5 y6 b7 b8 y9 b9
D C N A M+16 A F C AA C F A M+16 A N C D
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000
1.217,45 AMU, +2 H (Parent Error: -7,4 ppm)
12
Phosphatidylinositol-4-phosphate 5-kinase type-1 beta OS=Homo sapiens GN=PIP5K1B PE=1 SV=2
y3-NH3y6+1
parent+2H-H2O
y1
b2
y2 b3 y3
y4
b4 b5
y5
b6
y6
b7y7
I A D F G L A RR A L G F D A I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
861,47 AMU, +2 H (Parent Error: -12 ppm)
BCAP_HUMAN
a13-H2O+2Hy13+2H
b13+2H
b13+2H+1y7-NH3-H2Oa7-H2O+1y7-H2O
y7-H2O+1
b7-H2O-H2O
b7-NH3-H2O
a14-H2O+2H
a14-H2O+2H+1
y14-H2O+2H
y14-H2O+2H+1
b14-H2O-H2O+2H
a7
a7+1
y7+1
y8-H2O+1
b8-NH3-NH3
b8-NH3+1b16+2Ha17-H2O+2Ha17-H2O+2H+1
a19-NH3-NH3+2Hb19+2Ha20-H2O+2H+1
b5
y6
b6
y7
y8y9
b9y10 b11
L L T E G Q S F S SG L A E E D L S S F
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200
2.309,12 AMU, +3 H (Parent Error: 0,61 ppm)
RAC-alpha serine/threonine-protein kinase OS=Homo sapiens GN=AKT1 PE=1 SV=2
b9-H2O-H2Ob23-NH3-NH3+2Hb23-H2O+2H
b23-H2O+2H+1
b14-H2O-64
y25-NH3-H2O+2H b18+1
b18+2
y7
y8 b12y10 b13y11 y12 b15y13 b17y16b18
S L L S V S S G M+16 E G D N E D N E V P E V T R S RR S R T V E P V E N D E N D G E M+16 G S S V S L L S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 500 1000 1500 2000 2500
2.722,24 AMU, +2 H (Parent Error: 3,2 ppm)
14
HIPK3_HUMAN
y15+2H
y15+2H+1
b2y3b3 b4 b5 y5 b6 y6b7
y8 y10
Y G A E I V S A L D YE S H L Y D L A S V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000 1200
1.940,95 AMU, +3 H (Parent Error: -5,7 ppm)
Diacylglycerol kinase iota OS=Homo sapiens GN=DGKI PE=1 SV=1
y11-H2O-H2O+2Hb11-H2O+2H
y11-H2O+2H
a6-H2Ob6-H2Ob6-H2O+1b6-H2O+2
a12-H2O-H2O+2H
a12-NH3-H2O+2H
a12-NH3-NH3+2Ha12-H2O+2H
b6+1
y12-H2O-H2O+2H
b12-NH3-H2O+2H
y6-NH3-NH3b12-H2O+2H
a9+1
a9+2
parent+2H-H2O
y2 b5y4
y5
b6
y6 b8
y8
b9
b10
D M S H S G W R LL R W G S H S M D T
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000 1200
1.445,66 AMU, +2 H (Parent Error: -11 ppm)
C-Jun-amino-terminal kinase-interacting protein 4 OS=Homo sapiens GN=SPAG9 PE=1 SV=4
b16+2Hb9-H2O+1
a17-H2O+2Hb9+1
a10-H2O-64
b17-H2O-H2O+2H
y17-NH3-NH3+2H
parent+2H-NH3-64parent+2H-64
parent+2H+1-64
parent+2H+3-64
parent+2H-H2O
b7 y6
b8
y7
b9
y8 y9
A F S D FL F D S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
700 800 900 1000 1100
1.968,91 AMU, +2 H (Parent Error: -8,4 ppm)
15
LYN_HUMAN
b11+2H
y11+2Hy11+2H+1
a6-H2O
y9-H2O-H2O
b3 y3 b4y4b5y5 b6 b7 y8 y9
D F L S E E TS A L S A E T
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000
1.927,93 AMU, +3 H (Parent Error: -4,6 ppm)
MAK_HUMAN
a5
b6-H2O
b6-H2O+1b6-NH3+1
b11-NH3-H2O+2H
y5-H2Oy5-NH3
y12-H2O-H2O+2Hb12-NH3-H2O+2H
b8-NH3+1
b8-NH3+2
b9-NH3+1
b9-NH3+2
b2 b3 y3b4b5
y4
b6
y5
b7
y6 y7
y9
y10
y11
L N T L E T E E W F F KK F F W E E T E L T N L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.555,75 AMU, +2 H (Parent Error: -10 ppm)
AKAP1_HUMAN
a8-H2O-H2Oy8-H2Oa16-H2O-H2O+2Ha16-H2O+2Ha16-NH3+2H+1
y8+1
a16+2H+1
y3 b4 y5 y6y7
b7
y8
y9 y10b11
b12 y12b13 b14
S D W P E G Y Q L A S S M N F RR F N M S S A L Q Y G E P W D S
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750
1.886,83 AMU, +2 H (Parent Error: 0,93 ppm)
16
SRPK3_HUMAN
b13-H2O-64+2H
y10+1
y5 y6b6 b7
y7b8y8 b9
y10
b11
L I D F G I A N Q M+16 Q P D T T S V V KK V V S T T D P Q M+16 Q N A I G F D I L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750 2000
2.092,04 AMU, +3 H (Parent Error: -9,1 ppm)
CAMKV_HUMAN
b11+2Hy13+2H
a12-H2O+2H
y7+1
b12-NH3-NH3+2H
a14+2H+1
b15+2H
b15+2H+1
b18+2H
y3 y4
b3
y6
y7
b6
y8
b7 b9y10
b10
V H S+42 E N+14 N+14 A C I NG T S S T K F N I C A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000 1200
2.162,04 AMU, +3 H (Parent Error: -8,4 ppm)
KAP0_HUMAN
b11+2Hy13+2H
a12-H2O+2H
y7+1
b12-NH3-NH3+2H
a14+2H+1
b15+2H
b15+2H+1
b18+2H
y3 y4
b3
y6
y7
b6
y8
b7 b9y10
b10
V H S+42 E N+14 N+14 A C I NT S S T K F N I C A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000 1200
2.162,04 AMU, +3 H (Parent Error: -8,4 ppm)
17
KSR2_HUMAN
b7-H2Oa15+2H+1
b8+1a9-H2O-H2Ob9-H2O
a17-H2O-H2O+2Hb9+1
b9+2
y8-H2O-H2O
b10-H2O
y5 b7y6
b8
y7
b9
y8
y9
b11
y10
y11
y12 y13b14y14 b16
L T V A D A L E P V Q F E D G Q KK Q G D E F Q V P E L A D A V T L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750
1.858,93 AMU, +2 H (Parent Error: -7,5 ppm)
M3K3_HUMAN
b8-H2O+2H
y9-H2O-H2O+2H
a5
parent+2H-H2O
y1 b3 y3b4
y4
b5 y5b6
y6 b7 y7b8
E V M A Y R Q T RR T Q R Y A M V E
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.152,56 AMU, +2 H (Parent Error: -13 ppm)
Dual specificity protein kinase TTK OS=Homo sapiens GN=TTK PE=1 SV=2
b13-H2O-64+2H
y10+1
y5 y6 b6 b7
y7b8y8 b9
y10
b11
F G I A N Q M+16 Q PS T T D P Q M+16 Q N
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200
2.092,04 AMU, +3 H (Parent Error: -9,1 ppm)
18
cGMP-dependent protein kinase 1 OS=Homo sapiens GN=PRKG1 PE=1 SV=3
b4-H2O
b9-H2O+2H
y10+2H
b11-NH3+2H+1
y1 b4 b5
y4
b6
y5 y6 b7 y7
L G K G D W F GR V D E G Q L A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800
2.105,05 AMU, +4 H (Parent Error: -4,5 ppm)
Mitogen-activated protein kinase associated protein 1 (Fragment) OS=Homo sapiens GN=MAPKAP1 PE=4 SV=1
y8+2H
b5-H2O-H2O
a10-NH3-H2O+2Ha5
b5-H2O
y10-H2O-H2O+2Hy5-H2O
b10-NH3+2H
y1
b2y2 y3 y4b4 y6 b6
b7 b8
y8
M T V V T M A S A RR A S A M T V V T M
m/z
Rela
tive
Inte
nsity
0,0%
5,0%
10,0%
15,0%
0 200 400 600 800 1000
1.065,53 AMU, +2 H (Parent Error: 0,43 ppm)
Adenosine kinase OS=Homo sapiens GN=ADK PE=1 SV=2
y18+2H
b9+1
y20-H2O-H2O+2H
y20-NH3-NH3+2Ha10
a10+1
y11+1
y12+1a12+1 b16+1
y3
y4y5
y6
y7 b7y8
b8
y9
b9
y10
y11
b11
y12
b12
y13
b13y14
b14y15b15
b16
b17 b18
V M P Y V D I L F G N E T E A A T F A RR A F T A A E T E N G F L I D V Y P M V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750 2000
2.243,09 AMU, +2 H (Parent Error: -5,4 ppm)
19
Rho-associated protein kinase 1
y8+2H
b5-H2O-H2O
a10-NH3-H2O+2Ha5
b5-H2O
y10-H2O-H2O+2Hy5-H2O
b10-NH3+2H
y1
b2y2 y3 y4b4 y6 b6
b7 b8
y8
M T V V T M A S A RR A S A M T V V T M
m/z
Rela
tive
Inte
nsity
0,0%
5,0%
10,0%
15,0%
0 200 400 600 800 1000
1.065,53 AMU, +2 H (Parent Error: 0,43 ppm)
cDNA FLJ75392, highly similar to Homo sapiens hexokinase II (HKII) mRNA OS=Homo sapiens PE=2 SV=1
b7+2Hb8-H2O-64+2H
a8+2H
y5+0
b2
y2
y3b3
y4
y5
b6
M+16 E R GV P A G A H T
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 300 400 500 600 700
1.954,00 AMU, +4 H (Parent Error: 2,7 ppm)
cDNA FLJ56606, highly similar to Serine/threonine-protein kinase TAO2 (EC 2.7.11.1) OS=Homo sapiens PE=2 SV=1
a8
a15-H2O+2H
b15-NH3-NH3+2H
a15+2H
a15+2H+1
y2 y3b5 y4b6 y5
y6
b8
y7b9
y8b10 b11 y11 y14
L L L L R N S P Q P L RL P Q P S N R L L L L A G G
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
250 500 750 1000 1250 1500
1.603,96 AMU, +2 H (Parent Error: 1,8 ppm)
20
Mitogen-activated protein kinase kinase kinase 4 OS=Homo sapiens GN=MAP3K4 PE=1 SV=2
y12+2Hb13-H2O+2H
b13+2H
b13+2H+1 a9-H2O
b3 y2 y3b4 y4 b5y5b6
y6b8 b9
D I W S L G C V V IK G T V M+16 E I V V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000 1200
2.063,06 AMU, +3 H (Parent Error: 3,2 ppm)
Serine/threonine-protein kinase PAK 6 OS=Homo sapiens GN=PAK6 PE=1 SV=1
y11-H2O+2H
a13-H2O-H2O+2H
y2b2 b3 b4 b5
y5
y7 b8 y9
P E I S A P Q N F QP D F S T H V R H
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800 1000 1200
2.490,25 AMU, +4 H (Parent Error: 0,22 ppm)
FGFR2_HUMAN
b5-H2Oy10-H2O+2H
b12+2H
b7-NH3+1
a13-H2O+2Ha13-H2O+2H+1
y13-H2O-H2O+2H
y13-NH3-H2O+2Hy13-NH3-NH3+2H
b13-H2O+2H+1
b10+1
y3
b6b7
b8 y7b9 y8
b10
P A N C T N E L Y M M M RR M M M Y L E N T C N A P
m/z
Rela
tive
Inte
nsity
0%
20%
40%
60%
0 250 500 750 1000 1250 1500
1.572,66 AMU, +2 H (Parent Error: 1,5 ppm)
21
TRPM6_HUMAN
b7+2Ha15-H2O+2H
b7-NH3-H2O
b7+1
b7+2
a16-H2O+2H
y16-H2O-H2O+2H
b16-NH3-NH3+2H
y4 y5 b5 b6y7 y8
b7
y11 b12
E K M E D V N C S CC S C N V D E M K E
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200 1400
2.017,80 AMU, +2 H (Parent Error: 2,6 ppm)
TBRG4_HUMAN
b9+2Hb5-H2O
y9+2H
a10-H2O+2H
b10-NH3-H2O+2H
b10-H2O+2H
y1 b2 y2b3
y3
b4
y4b5
y5
b6y6
b7
y7
b8y8 b9
y9
E L Q S V E Q E V RR V E Q E V S Q L E
m/z
Rela
tive
Inte
nsity
0%
20%
40%
0 200 400 600 800 1000 1200
1.215,60 AMU, +2 H (Parent Error: -12 ppm)
OBSCN_HUMAN
b14-H2O+2Hb8-H2O-H2Oa8
a15-H2O+2H
y6+1
b3b4
y4b6
y5b7
y6
y7 b10 y8
G G S S S S S S S SF P A L E N D
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800
2.057,87 AMU, +3 H (Parent Error: -9,1 ppm)
22
TRIO_HUMAN
b8-128
a7-NH3+1
a16-64+2Hb16-H2O-64+2Hb16-64+2Hb8-H2O+1
b8-NH3+1
b8-NH3+2
a16-H2O+2Ha16-H2O+2H+1
a16-NH3+2H+1
y16-NH3-H2O+2H
b16-NH3-H2O+2H
b16-NH3-NH3+2H
a16+2H
b4 y5
y6
y7 b7 b8y8 b9 y11
b12b14
M+16 N V G R L Q G F D GF G Q L R G V N M+16 M+16 D N
m/z
Rela
tive
Inte
nsity
0,0%
2,5%
5,0%
7,5%
10,0%
12,5%
500 750 1000 1250 1500
1.815,77 AMU, +2 H (Parent Error: -7,5 ppm)
PAPS1_HUMAN
y10-H2O-H2O+2Hb10-NH3-H2O+2H
b2b3
y3
b4
y4b5y5
b6
y6
b7y7
y8
b9
T D A E T L P A L KK L A P L T E A D T
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.057,56 AMU, +2 H (Parent Error: -9,4 ppm)
TRI27_HUMAN
b10+2Hy10+2H
b5-NH3-H2Oy5-H2Oa11+2Hb5-H2Ob5-H2O+0b5-NH3
y11-H2O+2Hy6-NH3-H2Oy11+2H+1
y6-H2O+1
a12-H2O-H2O+2H
y6+1
y6+2
y12-H2O-H2O+2H
y12-NH3-H2O+2H
b12-NH3-NH3+2H
a12+2H
b12-H2O+2H+1
b3y4b4
y5b5
y6
b6y7b7
y8b8
b9y9b10y10
L Q D I G D T L SS L T D G I D Q L L
m/z
Rela
tive
Inte
nsity
0%
20%
40%
60%
200 400 600 800 1000 1200
1.358,70 AMU, +2 H (Parent Error: -11 ppm)
23
Tribbles homolog 1 OS=Homo sapiens GN=TRIB1 PE=1 SV=2
a6-H2O
y2b2 b3 y4b4 y5 b5
y6
y7 b7y8 b8
y9
b10 b11 b12
L L D A D D A A A V A A KK A A V A A A D D A D L L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.242,65 AMU, +2 H (Parent Error: 2,0 ppm)
KALRN_HUMAN
b8-H2O+2H+0b8-H2O+2H
b8+2H+0
b8+2H
y1b2 y2 b3 y3 b4 y4 b5 y5
b6
y6 b7
N H D R I RL D E Q R I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
200 400 600 800
1.537,77 AMU, +3 H (Parent Error: -6,3 ppm)
EPHA7_HUMAN
671,1?676,9?
679,6?
934,5?975,5?
b10-H2O-H2O+2Hb12-NH3-H2O+2H
y6-H2O-H2Oy13+2H
a9
b2
b4
y4
b5
b6y5 b7y6 b8y8b9
y9
b10y10b11
I D T I A A D E S F T Q G D L G E R KK R E G L D G Q T F S E D A A I T D I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750 2000
2.065,00 AMU, +3 H (Parent Error: 2,5 ppm)
AAK1_HUMAN
25
y12+2H
y10+1
y2 b3y3
b4y4
b5
y5 b6y6 b7y7
b8y8
b9y9 b10
y10
b11y11b12y12
b13 y13
L T D P I P T T E T S I A P RR P A I S T E T T P I P D T L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.610,84 AMU, +2 H (Parent Error: -10,0 ppm)
STK4_HUMAN
479,4?500,2?y10-NH3-H2O+2H
b10-H2O+2H
parent+2H-H2O
b2y1b3 y2 b4 y3
y4
b5y5
b6 b7
y6
b8y7
b9
y8
A V G D E M G T V RR V T G M E D G V A
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.033,48 AMU, +2 H (Parent Error: -14 ppm)
COASY_HUMAN
533,4?
542,6?
a5-H2O-H2O
a11-H2O+2Ha11-H2O+2H+0
y11-H2O-H2O+2H
b11-H2O-H2O+2H
parent+2H-H2O
parent+2H-NH3
y1y2
b3 b4
y3
y4
b5
y5
b6
y6
b7b8y7
y8
b9
b10
D G L S E A A A Q S RR S Q A A A E S L G D
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.103,52 AMU, +2 H (Parent Error: -9,4 ppm)
26
TBRG4_HUMAN
591,1?
y9+2H
b10-NH3-H2O+2H
b10-H2O+2H
parent+2H-H2O
y1 b2 y2b3
y3b4
y4b5
y5
b6 y6 b7
y7
b8 y8 b9y9
E L Q S V E Q E V RR V E Q E V S Q L E
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.215,60 AMU, +2 H (Parent Error: -12 ppm)
TRIB1_HUMAN
a6-H2O
y2b2 b3 y4b4 y5 b5
y6
y7 b7y8 b8
y9
b10 b11 b12
L L D A D D A A A V A A KK A A V A A A D D A D L L
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.242,65 AMU, +2 H (Parent Error: 2,0 ppm)
PAK2_HUMAN
553,1?
a9-H2O+2H
y9-H2O-H2O+2Hy9-NH3-H2O+2H
parent+2H-H2O
parent+2H-NH3
b2 y2 y3 b4
y4
b5 y5 b6 y6 b7
y7
b8
y8
M T D E E I M E KK E M I E E D T M
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000
1.124,47 AMU, +2 H (Parent Error: -11 ppm)
27
KAPCA_HUMAN
785,2?
794,1?
806,4?812,4?821,1?
996,5?
997,5?a9-H2O+2H a6-H2Oa13+2H+1
b14-NH3-H2O+2H
y7-H2O+1
b9+1parent+2H-H2O
y3y4
b5y5
b6
y6
b7b8 b9y9 b11
y11b12y12
F P S H F S S D L K D L L RR L L D K L D S S F H S P F
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500
1.660,86 AMU, +2 H (Parent Error: 2,2 ppm)
NTKL_HUMAN
973,5?
internal PELEQYD
y17+2Hy17+2H+1
internal PELEQYDPy18+2H+1
y10-H2O
a9
y19-NH3-H2O+2H
parent+2H-NH3
y4
b5y5
y6 b6
y7
b7
y8 b8
b9 y11
b10
b11y12
b12 y13b13 b14y14b15y15
G I P E L E Q Y D P P E L A D S S G RR G S S D A L E P P D Y Q E L E P I G
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 250 500 750 1000 1250 1500 1750 2000
2.071,96 AMU, +2 H (Parent Error: -7,2 ppm)
MK08_HUMAN
y1
b2
y2 b3y3y4 b4
b5
y5
b6
y6
b7y7
I L D F G L A RR A L G F D L I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
903,51 AMU, +2 H (Parent Error: -13 ppm)
28
SPS1_HUMAN
684,5?
685,4?
693,5?
694,5?
a12-H2O+2H
y5 b4b5
b6y7 y8
b7 y9b8 b9 y10 b11 y13
V A P Q V A T Q N VP T P N V N Q T A V
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
400 600 800 1000 1200
2.106,09 AMU, +3 H (Parent Error: -7,8 ppm)
KS6B2_HUMAN
490,3?
491,2?587,4?
y9+2H
y12-H2O+2H
y5+0
a8
parent+2H-H2O
y1 b4y2
y3
b6
y4
b7
y5
b8
y6
b9
y7 b10y8
b11
y9
y10b12y11 y12
I G G G P G D A A D V Q RR Q V D A A D G P G G G I
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800 1000 1200
1.211,58 AMU, +2 H (Parent Error: -12 ppm)
HCK_HUMAN
350,8?
parent+2H-H2Oparent+2H-NH3
y1b2
y2b3
y3
b4
y4
b5
y5
b6
D A W E I P RR P I E W A D
m/z
Rela
tive
Inte
nsity
0%
25%
50%
75%
100%
0 200 400 600 800
885,43 AMU, +2 H (Parent Error: -14 ppm)
29