Supplementary table I: Sequences and masses of Bd2400 peptides identified by LC-MS/MS. Masses include modifications on cysteine or methionine residues. Spots from the 2D analysis of the extracellular fraction were picked and grouped together in eleven groups according to their molecular weight (MW) and their intensity. Each group was submitted individually to in-gel proteolysis and liquid chromatography tandem mass spectrometry (LC-MS/MS) that were performed at theSmolerProteomicsCenter of the Technion, Israel. Proteins were in-gel digested with trypsin and dehydrated with acetonitrile, then rehydrated with 10% acetonitrile in10 mM ammonium bicarbonate containing trypsin [modified trypsin (Promega) at a 1:100 enzyme-to-substrate ratio]. Gel pieces were incubated overnight at 37oC and the resulting peptides were recovered and analyzed. Tryptic peptides were resolved by reverse-phase chromatography on 0.1 X 200-mm fused silica capillaries (J&W, 100 micrometer ID) packed with Everest reversed phase material (Grace Vydac). The peptides were eluted with linear 50 minutes gradients of 5 to 95% of acetonitrile with 0.1% formic acid in water at flow rates of 0.4 µl per min. Mass spectrometry was performed by an ion-trap MS (LCQ-DecaXP, Finnigan) in a positive mode using repetitively full MS scan followed by collision induces dissociation (CID) of the three most dominant ions selected from the first MS scan. The MS data were clustered and analyzed using the Pep-Miner searching the bacteria database (Beer et al. 2004).
Mass / Sequence705.3135 / AC(+57)GVNGK
905.494 / TGVITGETK
928.4412 / EFLTSFAN
988.4191 / LC(+57)DANPEAV
995.4473 / IFVC(+57)VDDK
995.4653 / IFVC(+57)VDDK
1002.537 / LNWVEVSR
1000.556 / C(+57)QNELPVIK
1144.52 / LC(+57)DANPEAVR
1149.499 / SEEVAC(+57)GVNGK
1248.568 / VSEEVAC(+57)GVNGK
1265.649 / LPDGSLLAHGASQ
1330.603 / C(+57)ADQVKPVEER
1330.621 / C(+57)ADQVKPVEER
1364.667 / ISGPQAGFVFNPM
1400.648 / TVEHSFFNVGTY
1428.669 / PVDPNAC(+57)TVAGATR
1477.801 / LPDGSLLAHGASQVL
1594.688 / TVIDDMDC(+57)LEDLR
1610.688 / TVIDDM(+16)DC(+57)LEDLR
1614.61 / PTDNFTAC(+57)TADC(+57)GGK
1640.864 / LPDGSLLAHGASQVLY
1661.73 / WNFGDGGTAANQSVTH
1685.647 / APTDNFTAC(+57)TADC(+57)GGK
1779.853 / VIEDLDVDPEPEVPVD
1824.858 / IFVC(+57)VDDKGVTVDDSR
1841.939 / LPDGSLLAHGASQVLYSN
1848.71 / YAPTDNFTAC(+57)TADC(+57)GGK
1876.932 / IATMM(+16)HVYLPAC(+57)LVNK
1876.932 / IATM(+16)MHVYLPAC(+57)LVNK
1879.93 / LGDNQEPWVTLDHSIR
1891.849 / ESQGELVSEEVAC(+57)GVNGK
1892.932 / IATM(+16)M(+16)HVYLPAC(+57)LVNK
2024.912 / VTVMTVIDDMDC(+57)LEDLR
2040.912 / VTVM(+16)TVIDDMDC(+57)LEDLR
2614.242 / TVEHSFFNVGTYQVQATVTDADGK
2791.356 / SQSSVTTIEEANSSATC(+57)PANQIGVIVK
2878.338 / VIEDLDVDPEPEVPVDPNAC(+57)TVAGATR