Biology Reference
In-Depth Information
0.4
687.81
m/z
MAPA
0.3
1398.72
606.37
m/z
MAPA
0.2
912.11
1059.55
650.39
699.9
734.38
m/z
MAPA
841.95
873.46
0.1
601.79
672.87
636.89
848.92
814.44
619.38
596.67
590.84
1032.56
744.82
565.82
982.03
1059.52
731.37
795.43
669.89
894.5
751.37
451.74
596.05
763.73
1087.59
128745
452.22
590.8
808.95
1270.06
828.43
488.24
734.89
514.28
976.48
842.37
153630
676.36
505.26
191582
196366
772.41
1067.55
803.97
173725
141319
523.28
965.95
469.74
701.34
443.77
898.5
685.39
573.3
677.34
54608
788.3
465.71
1075.53
740.39
781.35
816.45
1044.2
625.86
773.8
865.
1049.08
610.32
811.43
196044
182896
18029
152892
571.32
789.43
931.95
994.17
892.49
738.94
597.87
932.82
1015
1129.55
855.44
677.37
617.84
919.2
779.43
582.31
1109.58
1152.58
1257.68
702.41
823.43
728.47
808.45
537.75
596.65
729.43
766.91
814.12
872.43
571.83
954.45
57890
1099.67
1537.8
196500
120516
546.28
633.83
1074.52
1221.6
712.37
755.86
870.54
1000.53
404.2
856.01
936.98
24552
813.49
1019.54
1023.01
884.91
784.99
590.79
859.49
603.31
733.39
1219.57
983.47
1040.95
1476.71
1165.54
794.41
1205.11
651.69
847.82
761.4
532.77
196298
998.5
162449
136279
540.29
134235
108283
871.94
751.39
633.3
1021
531.81
966.95
692.89
596.83
185200
83064
78348
897.48
1115.1
737.88
576.98
102889
24120
643.31
727.72
887.41
138059
541.61
680.03
713.37
877.38
506.26
1080.55
1034.54
681.43
602.34
573.82
769.4
639.34
724.86
1230.64
604.89
732.44
133318
850.44
564.8
908.49
431.73
189877
871.97
843.86
1219.6
1050.58
774.05
652.35
684.35
651.35
571.96
537.8
1010.56
740.94
636.66
796.43
882.41
590.27
1126.56
205916
2426 195555
127317
173281
1087.58
1285.64
33411
726.42
665.33
625.31
193810
596.8
176233
782.43
664.84
195493
139269
722.9
577.55
778.44
135614
504.78
627.81
940.55
777.9
1200.63
678.11
735.85
1107.59
189186
137163
933.5
725.35
196431
194541
76146
732.47
135220
467.26
617.32
986.99
714.39
778.9
0
ATPv
MASS WESTERN
185915
126382
698.34
704.34
801.48
163824
845.39
811.93
130914
129019
756.85
979.97
521.26
865.91
574.32
446.75
865.46
195910
24459
799.87
648.85
990.5
624.85
666.88
650.84
546.3
591.25
609.38
627.32
1052.55
1496.79
rbcL
693.39
8086
196354
155280
1190.61
642.87
1044.63
1056.51
548.81
555.78
187891
869.98
679.36
522.27
186676
196833
891.49
458.75
751.44
687.86
835.97
127879
1427.78
104827
130316
153656
978.51
562.82
932.5
167973
1104.21
767.47
650.36
938.48
631.32
1006.54
196624
184810
136076
810.39
582.82
128726
196612
820.48
639.35
55838
808.94
912.53
915.07
602.85
968.96
136037
953.98
509.78
atpB
924.45
867.81
607.35
707.36
663.35
937.46
1152.59
1101.05
194461
10441
129025
132210
841.44
187367
831.95
613.81
828.33
874.56
1065.89
584.31
1051.05
617.86
708.89
529.3
558.29
684.33
1433.7
1252.68
653.86
846.47
814.48
120177
597.82
644.43
764.44
1254.69
645.34
132263
484.92
1000.44
629.32
916.84
1097.05
1006.08
926.99
443.2
608.81
625.76
603.33
465.72
872.55
671.06
881.95
598.33
101528
782.44
467.75
1405.2
19329
191662
1051.04
196443
943.56
543.78
585.79
194915
1356.66
845.78
886.96
832.43
490.28
675.36
668.36
701.03
861.43
809.41
744.4
666.37
816.36
647.32
638.32
1364.71
1078.07
1434.73
136854
1231.12
1250.1
893.57
894.47
1101.51
867.42
1047.53
645.4
1285.62
538.8
GAPDH cyt
MASS WESTERN
648.34
735.36
831.92
192398
697.84
462.26
822.46
140452
196305
640.34
616.29
593.96
1231.61
700.88
486.8
797.76
510.77
956.83
808.38
1073.53
576.32
775.43
-0.1
196570
1000.52
566.77
557.28
1055
-0.2
m/z
MAPA
m/z
MAPA
1119.55
610.33
-0.3
-0.2
-0.1
0
0.1
0.2
0.3
PC 1
e
Precursor ion:
m/z 1199.6112 (3+)
T
LADYFDVIGGTG
GGLLTAMITTPNENNRPFAAAK
y
31
2+
…
y
24
2+
2 Dalton m/z shift!
(1)
P
LADYFDVIGGTG
GGLLTAMITTPNENNRPFAAAK
[y
22
- 2xNH
3
]
2+
3
]
Precursor ion:
m/z 1198.2795 (3+)
(2)
Fig. 2
(continued) analysis (PCA) and sample pattern recognition for a MASS WESTERN protein data matrix of
two growth conditions of
Chlamydomonas reinhardtii
—mixotrophic and photoautotrophic [
16
]. Visible is the
clear separation of the two growth types. The separation in the principal components analysis is due to the
differences in protein abundances identifi ed by proteotypic peptides. (
c
,
d
) MAPA and MASS WESTERN data
were combined using COVAIN. First, each single data matrix is uploaded in COVAIN. Then the different data sets
are combined in one data matrix by COVAIN-supported selection of samples versus variables alignments. This
new data matrix is z-transformed and analyzed by principal components analysis (PCA). All steps are per-
formed by COVAIN. The two growth conditions of
Chlamydomonas reinhardtii
—mixotrophic and photoautotro-
phic [
16
]—are clearly separated in (
c
). In the corresponding loadings plot of (
d
) either proteins (MASS WESTERN)
or tryptic peptides (MAPA) are assigned for sample classifi cation. This procedure leads to rapid identifi cation of
protein markers out of millions of variables [
3
]. (
e
) Identifi cation of a protein polymorphism using the MAPA
approach [
3
]. (
1
) De novo sequencing and detection of an amino acid exchange threonine versus proline. (
2
)
This specifi c protein is exclusively detected in a potato “Lady Claire” cultivar
