Table 1. Proteins Identified in MPF Spots.
| spot no. | protein name | locusa | signal peptideb | PSORTb localization (score)c | sera reactivityd | previously identified as immunoreactive and reference | previously identified as membrane (M), secreted (T), or surface (S) protein and reference |
|---|---|---|---|---|---|---|---|
| 1 | dihydrolipoamide dehydrogenase (Lpd)e | FTL_0311 | Cyto (9.97) | X | (21) | M21,37 | |
| ATP-dependent protease, ATP-binding subunit (HslU) | FTL_0964 | Cyto (9.97) | |||||
| glyceraldehyde-3-phosphate dehydrogenase (GapA)f,e | FTL_1146 | Cyto (9.97) | (38−40) | T35,41 | |||
| glutathione reductase (Gor)e | FTL_1248 | SpI | unknown | ||||
| 2 | elongation factor Tu (TufA)f | FTL_1751 | Cyto (9.97) | X, Y | (21,38,40,42−46) | M21 | |
| 3–4 | could not be identified with confidence | X | |||||
| 5 | outer membrane associated protein FopA1f | FTL_1328 | SpI | OM (9.93) | X | (21,38,40,42,45,46) | M,21,23,47,48 T,35 S25,28 |
| 6 | universal stress protein (Usp) | FTL_0166 | Cyto (9.97) | X | (39) | M37 | |
| outer membrane associated protein FopA1f | FTL_1328 | SpI | OM (9.93) | (21,38,40,42,45,46) | M,21,23,47,48 T,35 S25,28 | ||
| 7 | could not be identified with confidence | X | |||||
| 8 | outer membrane associated protein FopA1f | FTL_1328 | SpI | OM (9.93) | X, Y | (21,38,40,42,45,46) | M,21,23,47,48 T,35 S25,28 |
| 9–10 | could not be identified with confidence | X | |||||
| 11 | acetyl-CoA carboxylase, biotin carboxyl carrier protein subunit (AccB)f | FTL_1592 | unknown | X, Y | (21,39,40,43−46) | M21,47 | |
| 12 | acetyl-CoA carboxylase, biotin carboxyl carrier protein subunit (AccB)f | FTL_1592 | unknown | X | (39,40,43−46) | M21,47 | |
| 13 | hypothetical proteinf | FTL_0617 | Cyto (8.96) | X | (21,38,39,45) | M,21,37,49 T,35,41 S25 | |
| 50S ribosomal protein L9 (RplI)f | FTL_1026 | Cyto (9.97) | (38,40,45,46) | T35 | |||
| F0F1 ATP synthase subunit delta (AtpH) | FTL_1798 | Cyto (9.26) | M49 | ||||
| 14 | 50S ribosomal protein L7/L12 (RplL)f | FTL_1745 | unknown | X, Y | (21,38−40,42,46) | M21 | |
| 15 | F0F1 ATP synthase subunit epsilon (AtpC) | FTL_1794 | NC (SP) | unknown | X | M49 | |
| 16 | hypothetical protein | FTL_0105 | SpI | unknown | X, Y | (40,45,46) | M48 |
| 17 | 30S ribosomal protein S7 (RpsG) | FTL_0233 | Cyto (9.97) | X | |||
| 50S ribosomal protein L5 (RplE) | FTL_0248 | Cyto (9.97) | |||||
| 30S ribosomal protein S5 (RpsE)e | FTL_0253 | NC (SP) | Cyto (9.26) | (46) | |||
| 50S ribosomal protein L15 (RplO) | FTL_0255 | NC (SP) | Cyto (9.26) | ||||
| outer membrane protein OmpHf,e | FTL_0536 | SpI | unknown | (38,40) | M37,47 | ||
| (3R)-hydroxymyristoyl-ACP dehydratase (FabZ)e | FTL_0538 | Cyto (9.97) | |||||
| 50S ribosomal protein L13 (RplM) | FTL_1187 | NC (SP) | Cyto (9.26) | ||||
| 18 | hypothetical protein | FTL_0571 | SpII | unknown | X | (45) | M47 |
| 19 | could not be identified with confidence | X | |||||
| 20 | 30S ribosomal protein S7 (RpsG) | FTL_0233 | Cyto (9.97) | X | |||
| 50S ribosomal protein L16 (RplP) | FTL_0243 | Cyto (9.97) | |||||
| 50S ribosomal protein L5 (RplE) | FTL_0248 | Cyto (9.26) | |||||
| 30S ribosomal protein S8 (RpsH) | FTL_0250 | Cyto (9.26) | |||||
| 30S ribosomal protein S5 (RpsE)e | FTL_0253 | NC (SP) | Cyto (9.26) | (46) | |||
| 50S ribosomal protein L15 (RplO) | FTL_0255 | NC (SP) | Cyto (9.26) | ||||
| 50S ribosomal protein L17 (RplQ) | FTL_0262 | Cyto (9.97) | |||||
| hypothetical protein (annotated as a pseudogene)e | FTL_0349 | unknown | |||||
| peptide methionine sulfoxide reductase (MsrB)e | FTL_0379 | SpI | Cyto (9.26) | ||||
| lipoprotein (LpnA)f | FTL_0421 | SpII | OM (10.00) | (21,40,43,44,46) | M,21,23,47,49 S26 | ||
| 30S ribosomal protein S9 (RpsI) | FTL_1186 | NC (SP) | Cyto (9.97) | ||||
| 50S ribosomal protein L10 (RplJ) | FTL_1746 | Cyto (9.26) | M37 | ||||
| 50S ribosomal protein L11 (RplK)e | FTL_1748 | NC (SP) | Cyto (9.26) | T35 | |||
| Sau5/YciO/YrdC family proteine | FTL_1913 | unknown | |||||
| 21 | lipoprotein (LpnA)f | FTL_0421 | SpII | OM (10.00) | X | (21,40,43,44,46) | M,21,23,47,49 S26 |
| 22 | lipoprotein (LpnA)f | FTL_0421 | SpII | OM (10.00) | X | (21,40,43,44,46) | M,21,23,47,49 S26 |
| 50S ribosomal protein L10 (RplJ) | FTL_1746 | Cyto (9.26) | M37 | ||||
| 23 | 50S ribosomal protein L10 (RplJ) | FTL_0235 | Cyto (9.26) | X | |||
| 50S ribosomal protein L22 (RplV)e | FTL_0241 | Cyto (9.26) | |||||
| 50S ribosomal protein L24 (RplX) | FTL_0247 | Cyto (9.97) | |||||
| hypothetical proteinf | FTL_0617 | Cyto (8.96) | (21,38,39,45) | M,21,37,49 T,35,41 S25 | |||
| histone-like protein HU form B (HupB)e | FTL_0895 | NC (SP) | Cyto (9.26) | (21) | M,21 T35 | ||
| 24 | histone-like protein HU form B (HupB)e | FTL_0895 | NC (SP) | Cyto (9.26) | X | (21) | M,21 T35 |
| 50S ribosomal protein L31 (RpmE)e | FTL_1303 | NC (SP) | Cyto (9.26) | ||||
| 30S ribosomal protein S16 (RpsP)e | FTL_1738 | Cyto (9.26) | |||||
| 25 | pyruvate dehydrogenase, E1 component (AceE) | FTL_0309 | Cyto (9.97) | Y | (40,50) | M23,50 | |
| 26 | chitinase family 18 protein (ChiA)f,e | FTL_1521 | SpI | unknown | Y | (38,45,46) | M23 |
| 27 | peroxidase/catalase (KatG)f | FTL_1504 | NC (SP) | Cyto (9.26) | Y | (21,38−40,42,45,46,50) | M,21,23,47,50 T35,41 |
| 28 | chaperone protein DnaKf | FTL_1191 | NC (SP) | Cyto (9.97) | Y | (21,38−40,42−46,50) | M,21,23,50 T,35,41 S25 |
| 29 | dihydrolipoamide acetyltransferase (AceF) | FTL_0310 | NC (SP) | Cyto (9.97) | Y | (21,40,45,50) | M21,37,47,50 |
| outer membrane associated protein FopA1f | FTL_1328 | SpI | OM (9.93) | (21,38,40,42,45,46) | M,21,23,47,48 T,35 S25,28 | ||
| chaperonin GroELf | FTL_1714 | Cyto (9.97) | (21,38−40,42−46) | M,21,23,43 T,35,41 S25 | |||
| 30 | chaperonin GroELf | FTL_1714 | Cyto (9.97) | Y | (21,38−40,42−46) | M,21,23,43 T,35,41 S25 | |
| dihydrolipoamide succinyltransferase component of 2-oxoglutarate dehydrogenase complex (SucB) | FTL_1783 | Cyto (9.97) | (21,40,45,46,50) | M21,37,50 |
a
NCBI reference sequence identification codes matching each loci are listed in Table S2 in the Supporting Information.
b
Signal peptide prediction; SpI, signal peptide cleaved by signal peptidase I; SpII, signal peptide cleaved by signal peptidase II; NC (SP), nonclassical signal peptide.
c
Subcellular localization predicted with PSORTb. Peri, periplasmic; Cyto, cytosol; OM, outer membrane; CM, cytoplasmic membrane.
d
X and Y designate proteins were reactive to sera from mice immunized with MPF and F. tularensis LVS vaccination, respectively.
e
Protein was not identified in the multidimensional LC–MS/MS analysis of MPF.
f
Protein was identified on the surface of F. tularensis LVS in this study.