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Table 1 List of identified xylem sap proteins from Brassica napus. Identifications of xylem sap proteins from the 2-DE gel shown in Figure 2. Sequences were determined from fragmentation spectra obtained by ESI-Q-TOF tandem MS. The resulting partial amino acid sequences were used for database searches with a BLAST algorithm optimized for short sequences. The isobaric amino acids isoleucine (I) and leucine (L), although not distinguishable by MS, are displayed as specified in the corresponding database sequences. BLAST E-values were determined using all non-redundant peptide sequences from one protein spot for another BLAST search. Bold letters in the partial sequences indicate amino acids identical to the database entry. Results of N-terminal secretion sequence predictions are indicated in the right column (Y= secretion sequence predicted, N= no secretion sequence predicted), * indicates that the observed molecular mass was lower and ** indicates that the observed mass was higher than expected.

From: Analysis of xylem sap proteins from Brassica napus

Spot No Similarity to Organism Acc. No. Sequence identity E value expected Mr observed Mr aS
1 Ubiquitin Arabidopsis thaliana At2g47110 TITLEVESSDTIDNVK 16/16 (100%) 9.00E-08 8525 12064 N
2 Ubiquitin Arabidopsis thaliana At2g47110 TITLEVESSDTIDNVK 16/16 (100%) 9.00E-08 8525 11944 N
3 Glycine-rich cell wall protein Arabidopsis thaliana At4g30450 AGSSAGSFAGSR 12/12 (100%) 0.011 9530 13863 Y
4 Pathogenesis-related protein PR1a Brassica napus AAB09587 SPQDYVNAHNQAR 13/13 (100%) 7.00E-39 17532 18145 Y
     LAAYAQNYADR 11/11 (100%)     
     QDSPQDYVNAHNQAR 15/15 (100%)     
     GPVQWDGTLAAYAQNYADR 19/19 (100%)     
     AGSSADFSGVSAVNLWVNEK 20/20 (100%)     
5 Glycine-rich cell wall protein Oryza sativa BAD62284 GGGYGEGGGYGGG 15/16 (94%) 2.00E-10 14152 17822 Y
     GGGYWGGGYGGGYGGG      
6 Invertase/pectin methylesterase inhibitor Arabidopsis thaliana At1 g47960 GSSADTSGLALILVDK 15/16 (94%) 2.00E-32 22326 18297 Y
     GVIDAGVEAAV 10/11 (91%)     
     ETPDFNLCVACPLDSDPR 15/18 (83%)     
     GSSADTSGLALILVDKIK 17/18 (94%)     
     ADVPEAIECCSK 10/12 (83%)     
     FGEDGVIDAGVEAAV 14/15 (93%)     
7 Papain-type cysteine proteinase XCP2 Arabidopsis thaliana At4g35350 LDHGVAAVGYGSSK 14/14 (100%) 4.00E-18 39618 16,603* Y
     ALAHQPVSVAIEASGR 16/16 (100%)     
     GVDLDHGVAAVGYGSSK 16/17 (94%)     
8 Papain-type cysteine proteinase XCP2 Arabidopsis thaliana At4g35350 LDHGVAAVGYGSSK 14/14 (100%) 4.00E-18 39618 16,977* Y
     ALAHQPVSVAIEASGR 16/16 (100%)     
     GVDLDHGVAAVGYGSSK 16/17 (94%)     
9 Invertase/pectin methylesterase inhibitor Arabidopsis thaliana At1g47960 GSSADTSGLALILVDK 15/16 (94%) 9.00E-38 22326 18297 Y
     ETPDFNLCVACPLDSDPR 15/18 (83%)     
     GSSADTSGLALILVDKIK 17/18 (94%)     
     ALDECASR 6/8 (75%)     
     ADVPEAIECCSK 10/12 (83%)     
     FGEDGVIDAGVEAAV 14/15 (93%)     
10 Glycine-rich protein Arabidopsis thaliana At4g30460 GEGGGYGGGYGGGGD 14/15 (93%) 1.00E-04 13738 19195 Y
11 Aspartyl protease family protein Arabidopsis thaliana At5g07030 LPPSAIAFNPATGAGTIFDSGTVYTR 24/26 (92%) 3.00E-15 46919 21,384* Y
     VLIDLPNSR 7/9 (77%)     
12 Disease resistance response/ dirigent protein Arabidopsis thaliana At1g64160 VIFDDPVTLDK 8/12 (67%) 3.00E-13 20573 22711 Y
     GCLNIMGADL 9/10 (90%)     
     DLSVVGGTGDFFFSR 14/15 (93%)     
13 Disease resistance response/ dirigent protein Arabidopsis thaliana At4g23690 VIFDDPVTLDQNYLSKPVSR 17/20 (85%) 1.00E-11 21412 24544 Y
     SVVGGTGDFFMSR 13/13 (100%)     
14 Endochitinase class I, putative Arabidopsis thaliana At2g43620 RDTIANVVTLSVFNSIFSK 12/19 (63%) 2.00E-08 30378 25856 Y
     QAFISAAAQSSDAYK 13/15 (87%)     
     FNGLPLLTDPDMVSR 13/15 (87%)     
15 Endochitinase class I, putative Arabidopsis thaliana At2g43620 RDTIANVVTLSVFNSIFSK 12/19 (63%) 2.00E-08 30378 25969 Y
     QAFISAAAQSSDAYK 13/15 (87%)     
     FNGLPLLTDPDMVSR 13/15 (87%)     
16 Endochitinase class I, putative Arabidopsis thaliana At2g43620 RDTIANVVTLSVFNSIFSK 12/19 (63%) 7.00E-13 30378 25744 Y
     QAFISAAAQSSDAYK 13/15 (87%)     
     FNGLPLLTDPDMVSR 13/15 (87%)     
     PVLDQGFGATTR 12/12 (100%)     
17 Germin-like protein, subfamily 2 Arabidopsis thaliana At1 g02335 YDPDALQDLCVAADK 14/15 (93%) 8.00E-07 23441 26543 Y
     KIPGLNTLSV 10/10 (100%)     
     GEVFVFPR 8/8 (100%)     
18 Thaumatin-like protein/ osmotin Brassica rapa AAN23104 RAPNTLAEYALK 12/12 (100%) 4.00E-04 incomplete 32060  
19 Gamma-glutamyltranspeptidase 1 Arabidopsis thaliana At4g39640 PPPAPANFIRPGK 13/13 (100%) 4.00E-D5 61190 26,312* Y
     VPPPAPANFIRPGK 14/14 (100%)     
     QFIVQES 7/7 (100%)     
20 Endochitinase class I, putative Arabidopsis thaliana At2g43620 TIANVVTLSVFNSIFSK 10/17 (59%) 2.00E-20 30378 28613 Y
     QAFISAAAQSSDAYK 13/15 (87%)     
     FLGLPLLTDPDFVAR 12/15 (80%)     
     IQITSDNYNYGAAGK 12/17 (71%)     
21 Xyloglucan:xyloglucosyl transferase Arabidopsis thaliana At5g57560 GQITNDGELLTLSLDK 14/16 (88%) 3.00E-36 32093 29928 Y
     LVPGNSAGTVTTLYLK 16/16 (100%)     
     WFDPTWFHTYTILSDNPQR 16/19 (84%)     
     IFTVDGTPIR 10/10 (100%)     
     NFESVGTLFPNNKPFR 13/16 (81%)     
22 Xyloglucan:xyloglucosyl transferase Arabidopsis thaliana At5g57560 GQITNDGELLTLSLDK 14/16 (88%) 3.00E-36 32093 30200 Y
     LVPGNSAGTVTTLYLK 16/16 (100%)     
     WFDPTVVFHTYTILSDNPQR 16/19 (84%)     
     IFTVDGTPIR 10/10 (100%)     
     NFESVGTLFPNNKPFR 13/16 (81%)     
23 Chitinase class IV, putative Arabidopsis thaliana At2g43570 PSFGSSISK 8/9 (89%) 2.00E-05 29775 31326 Y
     AAANSYPSFGSSISK 14/15 (93%)     
     LNENLLASPEK 10/11 (91%)     
     VAQDPPSAFK 7/10 (70%)     
24 Peroxidase 25 precursor Arabidopsis thaliana At2g41480 STVESHFDSDPTISPGLLR 19/19 (100%) 4.00E-09 35886 32665 Y
     EASNLPSPLDSVAVQK 16/16 (100%)     
     VALDIGSPSNFDVSFFK 15/17 (88%)     
25 Legume lectin family protein Arabidopsis thaliana At3g16530 LAVEFDTFQNK 10/11 (91%) 4.00E-06 30509 32360 Y
     GYWVQTR 7/7 (100%)     
Spot No Similarity to Organism Acc. No. Sequence identity E value expected Mr observed Mr aS
26 Legume lectin family protein Arabidopsis thaliana At3g15356 GDNLFFLGDAELG 11/13(85%) 7.00E-15 29749 32819 Y
     EFDTFKNK 7/8 (88%)     
     HVGININSMTSNVAEK 15/16 (94%)     
     TITIAPENVK 10/10 (100%)     
27 Chitinase class I Brassica napus CAA43708 DSFINAANTFPNFANSVTR 19/19 (100%) 5.00E-11 28733 30200 Y
     NGGNSGAVNAR 11/11 (100%)     
28 Legume lectin family protein Arabidopsis thaliana At3g16530 LAVEFDTFQNK 9/10 (90%) 5.00E-06 30509 31763 Y
     AGYWVQTR 8/8 (100%)     
29 Chitinase class I, putative Arabidopsis thaliana At4g01700 GFYPYEAFVEATR 13/13 (100%) 1.00E-08 31464 32819 Y
     GPIQLSWNYNYGQAGR 16/16 (100%)     
30 Peroxidase, putative Arabidopsis thaliana At4g36430 PGTVSCAADLLTLAAR 14/16 (88%) 4.00E-05 36164 34420 Y
     QNLSVLDIVSAAK 10/13 (77%)     
     GNISPLTGSSGEIR 14/14 (100%)     
31 Peroxidase, putative Arabidopsis thaliana At4g36430 DSSVLTGGPCWVVP 13/14 (93%) 7.00E-18 36164 36508 Y
     QGLDITDLVALSGSHTIGFSR 21/21 (100%)     
     NSDQVLFSS 9/9 (100%)     
     GNISPLTGSSGEIR 14/14 (100%)     
32 Peroxidase, putative Arabidopsis thaliana At1g49570 EAVVLTGGPFMAPVPLGR 16/18 (89%) 9.00E-06 38030 41164 Y
     GQPDPNLAASSDLLSK 15/16 (94%)     
     GNIGVFTGSDGVIR 13/14 (93%)     
33 Peroxidase ATP5a Arabidopsis thaliana At1g49570 EAVVLTGGPFMAPVPLGR 16/18 (89%) 7.00E-05 38030 40065 Y
     GNIGVFTGSDGVIR 13/14 (93%)     
  Peroxidase 59 precursor Arabidopsis thaliana At5g19890 LVEAYSQSQSLFFR 13/14 (93%) 6.00E-05 35023 40065 Y
34 Serine carboxypeptidase S10 family protein Arabidopsis thaliana At3g17180 GFIVGNPLTDDEYDNK 16/16 (100%) 1.00E-08 54145 40,498* Y
35 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78830 PLVYGGDSPNHDFR 11/14 (79%) 2.00E-09 50343 39,640 Y
     VVNNDEFGDYITEYDASYR 16/19 (84%)     
     GVTGFEILPNGNIVLHDK 17/18 (94%)     
     TGQPLVYGGDSPNHDFR 13/17 (76%)     
36 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78830 PLVYGGDSPNHDFR 11/14 (79%) 2.00E-09 50343 36,326* Y
     VVNNDEFGDYITEYDASYR 16/19 (84%)     
     GVTGFEILPNGNIVLHDK 17/18 (94%)     
     TGQPLVYGGDSPNHDFR 13/17 (76%)     
37 Peroxidase, putative Arabidopsis thaliana At4g33420 PTLSAGLIR 8/9 (89%) 1.00E-17 35975 37062 Y
     NDFDNAYFNALQR 13/13 (100%)     
     SGVLFSDQTLFNSPFTR 15/17 (88%)     
38 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78830 PLVYGGDSPNHDFR 11/14 (79%) 3.00E-08 50343 37,631* Y
     VVNNDEFGDYITEYDASYR 16/19 (84%)     
     ILPNGNIVLHDK 12/1 (100%)     
     TGQPLVYGGDSPNHDFR 13/17 (76%)     
39 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78830 PLVYGGDSPNHDFR 11/14 (79%) 3.00E-08 50343 35,967* Y
     VVNNDEFGDYITEYDASYR 16/19 (84%)     
     ILPNGNIVLHDK 12/1 (100%)     
     TGQPLVYGGDSPNHDFR 13/17 (76%)     
40 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78830 PLVYGGDSPNHDFR 11/14 (79%) 3.00E-08 50343 37,824* Y
     VVNNDEFGDYITEYDASYR 16/19 (84%)     
     ILPNGNIVLHDK 12/1 (100%)     
     TGQPLVYGGDSPNHDFR 13/17 (76%)     
41 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78830 PLVYGGDSPNHDFR 11/14 (79%) 3.00E-08 50343 36,326* Y
     VVNNDEFGDYITEYDASYR 16/19 (84%)     
     ILPNGNIVLHDK 12/1 (100%)     
     TGQPLVYGGDSPNHDFR 13/17 (76%)     
42 Peroxidase 59 precursor Arabidopsis thaliana At5g19890 KNAIPNINSAR 10/11 (91%) 0.003 35023 40280 Y
     EAYSQSQSLFFR 12/12 (100%)     
43 Peroxidase, putative Arabidopsis thaliana At4g33420 PTLAAGLIR 9/9 (100%) 1.00E-09 35975 37824 Y
     NTAEKDSPANLSLR 14/14 (100%)     
     NDFDNAYFNALQR 13/13 (100%)     
44 Cyclase family protein Arabidopsis thaliana At4g34180 DYLSFAAFDESPATHK 15/16 (94%) 1.00E-28 28384 34586 Y
     GLDYLSFAAFDESPATHK 17/18 (94%)     
     EFDSSFSGFFTDGAK 13/15 (87%)     
     LIGLDYLSFAAFDESPATHK 19/20 (95%)     
     GRDIIPVEALK 11/11 (100%)     
45 Peroxidase 21 (ATP2a) Arabidopsis thaliana At2g37130 DASLLLETAR 10/10 (100%) 5.00E-18 36741 35614 Y
     PTLDPDYALYLK 11/12 (92%)     
     ADNGYFHEQFSR 12/12 (100%)     
     LLSETNPLTGDQGEIR 16/16 (100%)     
46 Peroxidase, putative Arabidopsis thaliana At4g33420 PTLAAGLIR 9/9 (100%) 8.00E-40 35975 38018 Y
     GFTPQDVVALSGAHTLGVAR 20/20 (100%)     
     LTTPDSSFDSSFVNTLTK 13/18 (72%)     
     FDNAYFNALQR 10/11 (91%)     
     SGVLFSDQTLFNTPATR 17/17 (100%)     
47 Peroxidase, putative Arabidopsis thaliana At4g33420 DNTAEKDSPANLSLR 15/15 (100%) 1.00E-25 35975 37062 Y
     GFTPQDVVALSGAHTLGVAR 20/20 (100%)     
     NDFDNAYFNALQR 12/13 (92%)     
     LFSDQTLFNTP 11/11 (100%)     
48 Peroxidase 21 (ATP2a) Arabidopsis thaliana At2g37130 DASLLLETAR 10/10 (100%) 8.00E-09 36741 36691 Y
     LLSETNPLTGDQGEIR 16/16 (100%)     
49 Beta-1,3-glucanase (BG3) Arabidopsis thaliana At3g57240 LLLDVPNPDLQR 11/12 (92%) 3.00E-1B 30712 36691 N
     YISVGNEVQPS 11/11 (100%)     
     FVLPAMQNIDR 10/11 (91%)     
     TYVNNLIQTVK 11/11 (100%)     
Spot No Similarity to Organism Acc. No. Sequence identity E value expected Mr observed Mr aS
50 Beta-1,3-glucanase (BG3) Arabidopsis thaliana At3g57240 LLLDVPNPDLQR 11/12(92%) 3.00E-18 30712 37250 N
     YISVGNEVQPS 11/11 (100%)     
     FVLPAMQNIDR 10/11 (91%)     
     TYVNNLIQTVK 11/11 (100%)     
51 Polygalacturonase inhibitor protein 2 Brassica napus AAM94869 NFLQFDLSR 9/9 (100%) 5.00E-05 37117 40280 Y
     LQEFDTYSYFHNK 13/13 (100%)     
52 Peroxidase 59 precursor Arabidopsis thaliana At5g19890 PYLVQIVR 7/8 (88%) 2.00E-06 35023 41850 Y
     GFEVVDTIK 8/9 (89%)     
     LVEAYSQSQSLFFR 13/14 (93%)     
53 Peroxidase 59 precursor Arabidopsis thaliana At5g19890 PYLVQIVR 7/8 (88%) 5.00E-04 35023 43793 Y
     LVEAYSQSQSLFFR 13/14 (93%)     
54 Peroxidase 59 precursor Arabidopsis thaliana At5g19890 PYLVQIVR 7/8 (88%) 2.00E-06 35023 43793 Y
     GFEVVDTIK 8/9 (89%)     
     LVEAYSQSQSLFFR 13/14 (93%)     
55 Subtilase family protein Arabidopsis thaliana At1g20160 AVASAYGTFPTTVIDSK 16/17 (94%) 1.00E-17 81480 56,025* Y
     SILKPDITAPGVAILAAWTG 19/20 (95%)     
     LTYQVTVSATA 10/11 (91%)     
56 Subtilase family protein Arabidopsis thaliana At1g20160 SPVYPLIHGK 10/10 (100%) 2.00E-05 81480 56,025* Y
     ASAYGTFPTTVIDSK 14/15 (93%)     
     VSVETPAYQIQVTPEK 13/16 (63%)     
     VFGALT 6/6 (100%)     
57 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78850 KVPVDEQFLVVNK 10/13 (77%) 8.00E-16 49051 50573 Y
     TDYSPIEYNPDVR 13/13 (100%)     
     ILENGNFVIYDSSVK 13/15 (87%)     
     SVNTDGPYSLVFEDK 12/15 (80%)     
     TPKPIAVREGYEFFSK 12/16 (75%)     
58 Polygalacturonase/ glycoside hydrolase 28 Arabidopsis thaliana At3g61490 DATLLAAQDLEEYPVLK 14/17 (82%) 1.00E-10 51939 56471 Y
     DEYGIAFGFPTK 10/12 (83%)     
     IALGSEFSGGIEDVR 14/15 (93%)     
59 Subtilase family protein Arabidopsis thaliana At1g20160 SPVYPLIHGK 10/10 (100%) 1.00E-10 81480 58,848* Y
     ASAYGTFPTTVIDSK 14/15 (93%)     
     SLHPTWCPSAIR 10/12 (83%)     
     VSVETPAYQIQVTPEK 13/16(81%)     
     LTYQVTVSAADDDV 10/14(71%)     
     VFGALT 6/6 (100%)     
60 Subtilase family protein Arabidopsis thaliana At1g20160 SPVYPLIHGK 10/10 (100%) 1.00E-10 81480 56,025* Y
     ASAYGTFPTTVIDSK 14/15 (93%)     
     SLHPTWCPSAIR 10/12 (83%)     
     VSVETPAYQIQVTPEK 13/16(81%)     
     LTYQVTVSAADDDV 10/14(71%)     
     VFGALT 6/6 (100%)     
61 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78850 KVPVDEQFLVVNK 10/13 (77%) 6.00E-11 49051 50573 Y
     TDYSPIEYNPDVR 13/13 (100%)     
     ILENGNFVIYDSSVK 13/15 (87%)     
     SVNTDGPYSLVFEDK 12/15 (80%)     
     TPKPIAVREGYEFFSK 12/16 (75%)     
62 Curculin-like (mannose-binding) lectin Arabidopsis thaliana At1g78850 KVPVDEQFLVVNK 10/13 (77%) 6.00E-11 49051 51999 Y
     TDYSPIEYNPDVR 13/13 (100%)     
     ILENGNFVIYDSSVK 13/15 (87%)     
     SVNTDGPYSLVFEDK 12/15 (80%)     
     TPKPIAVREGYEFFSK 12/16 (75%)     
63 Subtilase family protein Arabidopsis thaliana At1g20160 AVASAYGTFPTTVIDSK 16/17 (94%) 3.00E-07 81480 56,926* Y
     TVTNVGGDRAVYK 10/13 (77%)     
     VFGALT 6/6 (100%)     
64 Polygalacturonase/ glycoside hydrolase 28 Arabidopsis thaliana At3g61490 DEYGIAFGFPTK 10/12 (83%) 9.00E-08 51939 5786B Y
     IALGSEFSGGIEDVR 14/15 (93%)     
     WTLLAAQDLEEYPVLK 14/16 (88%)     
65 Subtilase family protein Arabidopsis thaliana At1g20160 ESDVVLGGSK 9/10 (90%) 5.00E-08 81480 81188 Y
     SPVYPLIHGK 10/10 (100%)     
     AVASAYGTFPTTVIDSK 16/17 (94%)     
     SLHPTWCPSAIR 10/12 (83%)     
     TVTNVGGDRAVYK 10/13 (77%)     
     LTYQVTVSA 8/9 (89%)     
66 Subtilase family protein Arabidopsis thaliana At1g20160 ESDVVLGGSK 9/10 (90%) 5.00E-08 81480 79849 Y
     SPVYPLIHGK 10/10 (100%)     
     AVASAYGTFPTTVIDSK 16/17 (94%)     
     SLHPTWCPSAIR 10/12 (83%)     
     TVTNVGGDRAVYK 10/13 (77%)     
     LTYQVTVSA 8/9 (89%)     
67 Subtilase family protein Arabidopsis thaliana At1g20160 ESDVVLGGSK 9/10 (90%) 6.00E-07 81480 81188 Y
     AVASAYGTFPTTVIDSK 16/17 (94%)     
     TVTNVGGDRAVYK 10/13 (77%)     
     LTYQVTVSA 8/9 (89%)     
     VFGALT 6/6 (100%)     
68 Cucumisin-like serine protease (ARA12) Arabidopsis thaliana At5g67360 VEGASLLGFASGTAR 14/15 (93%) 1.00E-06 79415 78587 Y
     DGVAIGAFAAFER 12/13 (92%)     
     PALAILGNGK 10/10 (100%)     
     LNYPSFAVNVDGSGAYK 16/17 (94%)     
     TVTFTVDSSK 10/10 (100%)     
69 Germin-like protein Arabidopsis thaliana At3g05950 FTSGLNIAGNTIN 13/13 (100%) 7.00E-05 24736 76,260** Y
     AFQLDVNVVR 9/10 (90%)