TABLE 2.
Immunoreactive proteins identified by MALDI-TOF MS
| Group | Band no.a | Sourceb | Accession no.c | Characterization of protein |
Peptide matches |
|||||
|---|---|---|---|---|---|---|---|---|---|---|
| Name | Descriptiond | Mass (kDa) | pI | No. matched | Coverage (%) | MOWSE scoree/database | ||||
| Virulence | 01 | S, O, V | YP_003229794 | FliC | Flagellin H11 monomer | 50.92 | 4.70 | 181 | 85 | 8,628/NCBInr |
| 02 | S | AAO17297 | EatA | Autotransporter | 147.61 | 6.23 | 9 | 9 | 272/NCBInr | |
| 03 | S | YP_001464349.1 | Antigen 43 | Autotransporter | 97.27 | 5.92 | 12 | 10 | Sangerf | |
| 50 | S, O, V | AAX13509.2 | EtpA | TpsA adhesin | 176.92 | 9.77 | 19 | 8 | 375/NCBInr | |
| 08 | O | AAC41414.1 | CfaA | Periplasmic chaperone | 27.2 | 9.79 | 9 | 43 | Sanger | |
| 09 | O | A7ZK44 | FimD2 | Fimbrial usher | 95.2 | 5.42 | 20 | 19 | Sanger | |
| 17, 36, 54 | O, W, S | P02971 | CfaB | Pilin subunit | 17.45 | 7.93 | 5 | 48 | 252/NCBInr | |
| 49 | O | ABV16486 | Antigen 43 | Autotransporter | 97.29 | 5.56 | 4 | 5 | 75/NCBInr | |
| 43 | V | AAB02982 | LT-B | Heat-labile toxin b subunit | 14.06 | 8.76 | 5 | 30 | 145/NCBInr | |
| 43 | V | ABM92275 | CexE | Secreted ETEC protein, unknown function | 12.63 | 9.32 | 3 | 26 | 154/NCBInr | |
| 51 | V | Q9XD84 | TibA | Autotransporter/adhesin | 101.05 | 5.58 | 9 | 10 | 294/NCBInr | |
| Molecular chaperone | 22 | S | AAR21890 | GroEL | Chaperonin | 57.28 | 4.84 | 19 | 61 | 1,386/NCBInr |
| 21 | S | NP_308041 | DnaK (Hsp70) | Chaperonin | 69.07 | 4.83 | 32 | 60 | 1,587/NCBInr | |
| 06 | O | NP_285857 | (DegP/HtrA) | Membrane-associated serine protease | 46.80 | 7.88 | 17 | 52 | 548/NCBInr | |
| Outer membrane protein | 07, 20, 12 | O | NP_309068.1 | OmpA | Porin | 37.18 | 5.99 | 17 | 51 | 695/NCBInr |
| 39, 11 | V, O | AAB40749 | NmpC | Porin | 41.5 | 4.94 | 13 | 37 | 997/NCBInr | |
| 10 | O | YP_405685.1 | FecA homologue | Fe transport membrane receptor | 85.16 | 5.99 | 17 | 16 | Sanger | |
| 14, 42 | O, V | NP_752830 | OmpX | Ail-like adhesin? | 18.59 | 6.56 | 7 | 57 | 333/NCBInr | |
| 19 | O | NP_414692.1 | FhuA-like protein | Ferrichrome outer membrane transporter | 82.2 | 5.35 | 17 | 19 | Sanger | |
| 53 | O | NP_417795.1 | Bfr | Bacterioferrin | 18.48 | 4.69 | 8 | 67 | 472/NCBInr | |
| 59, 60 | O | YP_540467.1 | OmpW | Surface antigen 2 superfamily protein | 25.856 | 6.28 | 5 | 32 | 229/NCBInr | |
| Hypothetical | 34 | O | NP_288109.1 | Hypothetical | 7.89 | 9.6 | 2 | 75 | Sanger | |
| 45 | O | ZP_04533914.1 | Hypothetical | 75.53 | 8.91 | 20 | 27 | Sanger | ||
| 23 | S | NP_755083.1 | Hypothetical | 30.42 | 4.97 | 23 | 67 | 789/NCBInr | ||
| 28 | S | NP_311861.1 | Hypothetical | 26.41 | 9.35 | 10 | 53 | Sanger | ||
| 27 | S | YP_853622 | Hypothetical | Similar to APEC01 protein | 36.42 | 5.53 | 5 | 25 | 138/NCBInr | |
| 31 | S | YP_002408614 | Hypothetical | Conserved, hypothetical (phage origin) | 12.76 | 5.82 | 3 | 56 | 263/NCBInr | |
| 35 | W | NP_286070 | Hypothetical | Conserved hypothetical in EHEC strains | 9.92 | 5.36 | 3 | 49 | 72/NCBInr | |
| 15 | O | NP_288111 | Hypothetical | 11.58 | 4.83 | 4 | 45.7 | Sanger | ||
| 33 | ZP_04533918.1 | Hypothetical | Homologue in Crohn's isolate E. coli 3_2_53FAA | 16.65 | 10.10 | 9 | 44 | Sanger | ||
| 37 | V | YP_002388453.1 | YghJ | Predicted membrane lipoprotein | 168.65 | 4.92 | 13 | 13 | 624/NCBInr | |
| 41 | V | NP_288218 | Hypothetical | MipA domain; envelope biogenesis | 27.78 | 5.50 | 4 | 22 | 90/NCBInr | |
| 40 | V | YP_541993 | Hypothetical | Homologue in UPEC (UTI89) | 30.42 | 4.97 | 7 | 42 | 205/NCBInr | |
| Transcription/translation | 24 | S | NP_312218.1 | Elongation factor G | 77.53 | 5.24 | 22 | 37 | 791/NCBInr | |
| 30 | S | CAA47740.1 | H-NS | 15.53 | 5.43 | 5 | 45 | 363/NCBInr | ||
| Housekeeping/metabolic | 26 | S | H85928 | Phosphopyruvate hydratase | 45.57 | 5.32 | 19 | 54 | 644/NCBInr | |
| 44 | O | ZP_03072175.1 | Pyruvate dehydrogenase | 99.61 | 5.46 | 8 | 8 | 214/NCBInr | ||
| 29 | S | AAC43517 | Thiol peroxidase | 17.88 | 4.75 | 4 | 37 | 400/NCBInr | ||
| 32 | S | AAA24696 | Thioredoxin | 11.89 | 4.67 | 4 | 47 | 210/NCBInr | ||
| 25 | S | NP_285812.1 | Dihydrolipoamide dehydrogenase | 50.66 | 5.79 | 18 | 43 | 590/NCBInr | ||
The band number refers to the band from the stained gel. The bands were numbered chronologically in order of submission for MALDI-TOF MS identification.
Source: S, supernatant; O, outer membrane; V, vesicle; W, unfractionated whole-cell lysate.
The accession number refers to the closest homologue identified by BLAST(P) search.
TpsA, two-partner secretion exoprotein family; APEC, avian pathogenic Escherichia coli; EHEC, enterohemorrhagic Escherichia coli; UPEC, uropathogenic Escherichia coli.
MOWSE score generated using the MASCOT algorithm to query the NCBInr database.
Data for peptide lacking matches in the NCBInr database were matched to predicted proteins based on the Sanger sequence for strain H10407 using custom Perl scripts and FindPept.