Arabidopsis plants were treated with gaseous NO and S-nitrosylated proteins of leaves were enriched using the biotin switch method. Leaves of untreated plants served as control. Biotinylated proteins were affinity purified and analyzed by nanoLC/MS/MS after tryptic digestion. The MASCOT search engine was used to parse mass spectrometry data to identify proteins from primary sequence databases. The best-matching peptide identifying the protein is given. If there were further peptides found the number of the peptides is given. Hints confirming that the identified protein is a candidate for S-nitrosylation are given in the right column. The results of two separate experiments of each treatment were summarized in the table. Acc. No., accession number
Protein / Acc. No. / MW / NO/Identified peptide (score) / Control/Identified peptide (score) / Hints to S-nitrosylation
Metabolic enzymes
transketolase-like protein / T47886 / 81424 / SIITGELPAGWEK (61) +1 / TPGHPENFETPGIEVTTGPLGQGIANAVGLALAEK (72) +1
carbonic anhydrase, chloroplast precursor / NP_850490 / 29485 / EAVNVSLANLLTYPFVR (103) + 4
Carbonic anhydrase 2 / P42747 / 28326 / EAVNVSLANLLTYPFVR (103) +4 / EAVNVSLANLLTYPFVR (69) / (Ji et al., 1999; Klatt et al., 2000)
Glutamine synthase precursor, chloroplast / NP_198413 / 47381 / IIAEYIWIGGSGIDLR (37) +2 / (Motohashi et al., 2001)
malate dehydrogenase, cytosolic, putative2 / NP_17936 / 35548 / MELIDAAFPLLK (59) +1 / (Yamazaki et al., 2004)
probable malate dehydrogenase precursor, glyoxysomal / NP_196528 / 37346 / LLGVTTLDVAR (80) / ANTFVAEVLGLDPR (69)
malate dehydrogenase, mitochondrial / AAM64855 / 35754 / ALEGADLVIIPAGVPR (36) / (Foster and Stamler, 2004)
fructose bisphosphate aldolase, putative2 / AAK59548 / 42862 / TLLVSAPGLGQYISGAILFEETLYQSTTDGK (55) +2 / (Fratelli et al., 2002; Lind et al., 2002; Ito et al., 2003; Shenton and Grant, 2003)
probable triosephosphate isomerase1 / AAF70259 / 33325 / GGAFTGEISVEQLK (39) / (Fratelli et al., 2002; Ito et al., 2003; Shenton and Grant, 2003)
2- phosphoglycerate hydrolase (enolase) / NP_181192 / 47689 / VNQIGSVTESIEAVK (79) / (Fratelli et al., 2002; Lind et al., 2002; Shenton and Grant, 2003)
phosphoglycerate kinase–related1 / NP_187884 / 50081 / ELDYLVGAVSNPK (56) +4 / ADLNVPLDDNQTITDDTR (98) +5 / (Fratelli et al., 2002)
glyceraldehyde-3-phosphate dehydrogenase, cytosolic2 / JQ1287 / 36963 / GILGYTEDDVVSTDFVGDNR (83) +1 / (Mohr et al., 1996; Mohr et al., 1999; Klatt et al., 2000; Motohashi et al., 2001; Kuncewicz et al., 2003; Shenton and Grant, 2003)
ATP synthase CF1 alpha chain / NP_051044 / 55294 / LIESPAPGIISR (49) +2 / (Eaton et al., 2003)
ATP synthase CF1 beta chain / NP_051066 / 53900 / YKELQDIIAILGLDELSEEDR (96) +14 / GIYPAVDPLDSTSTMLQPR (108) +6
Redox-related proteins
peroxiredoxin - like protein1 / NP_190864 / 24669 / TVTVSSLTAGK (55) +1 / (Motohashi et al., 2001; Fratelli et al., 2002; Lind et al., 2002; Fratelli et al., 2003)
thioredoxin f1 / AAD35003 / 19339 / YDDLVAAIETAR (48) / (Ruelland and Miginiac-Maslow, 1999; Klatt et al., 2000; Motohashi et al., 2001)
Signalling/regulation proteins
Elongation factor 21 / A96602 / 94185 / STLTDSLVAAAGIIAQEVAGDVR (77) / (Fratelli et al., 2002; Shenton and Grant, 2003)
elongation factor Tu / NP_193769 / 51598 / QTELPFLLAVEDVFSITGR (53)
initiation factor 5A-4 related1 / NP_173985 / 17129 / VDYQLIDITEDGFVSLLTDSGGTK (59) / (Lind et al., 2002; Shenton and Grant, 2003)
Signalling/regulation proteins
putative RNA-binding protein / Q8LAU2 / 42576 / DLATAFLNVLGNEK (82) +1
SET domain transcriptional regulatory family / Q8VY03 / 57490 / KEGDDSEDLK (44)
50S ribosomal protein / AAM64725 / 19670 / IGSEISSLTLEEAR (40)
trigger factor-related / NP_200333 / 61695 / LLPGFLDAIIGIR (33)
Stress-related proteins
peptidylprolyl isomerase ROC41 / NP_191762 / 28190 / IVMGLFGEVVPK (37) / IVMGLFGEVVPK (39) / (Motohashi et al., 2001; Fratelli et al., 2002; Fratelli et al., 2003)
chaperonin 10 / T52122 / 26912 / ITPLPVSTGSTVLYSK (51) +1
chaperonin 60 beta precursor / JT0901 / 63792 / EVELEDPVENIGAK (57) / DLVGVLEDAIR (46) +1
cold-regulated protein (cor15a) / NP_181782 / 14596 / GDGNILDDLNEATK (44)
cold-regulated protein (cor15b) / NP_181781 / 14952 / SDGNILDDLNEATK (46)
heat shock cognate 70-1 / CAA54419 / 70028 / KIEDSIEQAIQWLEGNQLAEADEFEDK (64) / (Jaffrey et al., 2001; Fratelli et al., 2002; Lind et al., 2002; Fratelli et al., 2003; Shenton and Grant, 2003)
Proteins involved in photosynthetic processes
glyceraldehyde-3-phosphate dehydrogenase (NADP) A precursor, chloroplast / JQ1285 / 42481 / VVDLADIVANNWK (74) +1 / (Mohr et al., 1996; Mohr et al., 1999; Klatt et al., 2000; Motohashi et al., 2001; Kuncewicz et al., 2003; Shenton and Grant, 2003)
glyceraldehyde-3-phosphate dehydrogenase (NADP) B precursor, chloroplast / JQ1286 / 42769 / VPTPNVSVVDLVINVEK (62) +1 / VPTPNVSVVDLVINVEK (48) +2
riblose 1,5-bisphosphate carboxylase/oxygenase large chain / NP_051067 / 52922 / ESTLGFVDLLR (90) +6 / LSGGDHIHAGTVVGK (56) +2 / (Marcus et al., 2003)
ribulose bisphosphate carboxylase small chain 1a precursor / RKMUA1 / 20189 / IIGFDNTR (45) +1 / IIGFDNTR (58) +1 / (Motohashi et al., 2001)
ribulose bisphosphate carboxylase small chain 1b precursor / NP_198659 / 20273 / IIGFDNTR (58) +1
ribulose-bisphosphate carboxylase small chain B2 precursor / RKMUB2 / 20303 / IIGFDNTR (58) +1 / IIGFDNTR (58) +2
rubisco activase, large subunit / AAA20202 / 51948 / VPLILGIWGGK (34) / GLAYDTSDDQQDITR (81) +3 / (Zhang and Portis, 1999; Zhang et al., 2002)
photosystem I subunit II precursor / CAB52677 / 22335 / EAPVGFTPPQLDPNTPSPIFAGSTGGLLR (36)
Photosystem II P680 47 kDa protein / NP_051084 / 56001 / MPTFFETFPVVLVDGDGIVR (85)
Photosystem II D2 protein, fragment / AAO13251 / 35220 / NILLNEGIR (31) / (Carlberg et al., 1999)
photosystem II oxygen-evolving complex 23 / NP_172153 / 28078 / EIEYPGQVLR (37) +1 / KFVESAATSFSVA (54) +2
photosystem II oxygen-evolving complex 33 / NP_190651 / 34998 / NTAASVGEITLK (53) +4
probable photosystem II stability protein / AAK74049 / 44076 / VFLPIDPGVVLLDIAFVPDEPSR (36)
Rieske Fe-S protein / CAC03598 / 24334 / VLFVPWVETDFR (63)
Thylakoid lumenal 16.5 kDa protein, chloroplast precursor / O22773 / 25612 / WTTLTGLLEQLK (72)
Others
glycine decarboxylase complex H protein precursor–related / NP_181080 / 17936 / LTESPGLINSSPYEDGWMIK (48)
acyl carrier protein family / NP_194235 / 14536 / EQLALAADVPLTAESK (92) +1
endopeptidase Clp protease ATP-binding subunit / NP_566912 / 105706 / IASGDVPETIEGK (21) +1
putative alanine aminotransferase / AAK25905 / 53320 / TTILPAEDEMPEIMDSFK (37)
aminomethyltransferase / H86252 / 44417 / IGEITSGGFSPNLK (49) +1
probable major latex protein / AAK63869 / 17507 / VFDIIYEFIPK (37)
expressed protein / NP_564691 / 37387 / LNQVQAGLSALEEALK (37)
unknown protein / AAK59490 / 10554 / NPIDFVLGFMTK (43) / NPIDFVLGFMTK (58)
Literatur
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