TABLE 1.
Major cell wall-associated proteins identified in the mutanolysin extracts of GAS strains NS931, NS13, and S43 by MALDI-TOF peptide mass fingerprinting analysisa
Function or pathway | Spot | Protein | Accession no.b | Molecular mass (kDa)c | pIc | Peptide matchd | Coverage (%)e | Mutanolysin extract
|
Immunoreactive
|
Biotinylated
|
||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
NS931 | NS13 | S43 | NS931 | NS13 | S43 | NS931 | NS13 | S43 | ||||||||
Glycolysis | 4 | Putative pyruvate kinase | Q8K7A3 | 54.5 | 4.96 | 30 | 59.8 | + | + | + | + | + | + | + | + | |
5 | Putative NADP-dependent glyceraldehyde-3-phosphate dehydrogenase | Q8K707 | 49.5 | 4.96 | 19 | 56.1 | + | + | + | + | + | + | + | |||
11 | Phosphoglycerate kinase | Q8K5W7 | 42.0 | 4.86 | 23 | 70.0 | + | + | + | + | + | + | + | |||
17 | 6-Phosphofructokinase | Q8P0S6 | 35.8 | 5.34 | 22 | 50.7 | + | + | + | + | + | |||||
21 | Fructose-bisphosphate aldolase | P82486 | 31.1 | 4.87 | 13 | 50.3 | + | + | + | + | + | + | + | |||
27 | Triosephosphate isomerase | P82478 | 26.5 | 4.57 | 11 | 63.3 | + | + | + | + | + | + | + | + | ||
28 | 2,3-Bisphosphoglycerate-dependent phosphoglycerate mutase | Q8P0C1 | 26.0 | 5.10 | 19 | 77.1 | + | + | + | + | + | |||||
69 | l-Lactate dehydrogenase | Q99ZN5 | 35.1 | 5.14 | 9 | 30.7 | + | |||||||||
Carbohydrate metabolism | 1 | Putative transketolase | Q8K670 | 77.5 | 4.98 | 21 | 42.1 | + | + | + | + | + | + | + | + | + |
32 | Putative dTDP-4-keto-6-deoxyglucose-3,5-epimerase | Q9A046 | 22.4 | 5.07 | 10 | 42.1 | + | + | + | |||||||
41 | Putative lactoylglutathione lyase | Q9A121 | 14.4 | 5.09 | 4 | 30.4 | + | |||||||||
52 | Putative phosphoglucomutase | Q99ZH8 | 63.3 | 4.78 | 10 | 25.7 | + | |||||||||
54 | Putative phospho-sugar mutase | Q878L0 | 48.4 | 4.57 | 8 | 17.7 | + | |||||||||
60 | Putative dTDP-glucose-4,6-dehydratase | Q8P199 | 38.8 | 5.41 | 4 | 12.4 | + | + | ||||||||
61 | Glycerol-3-phosphate dehydrogenase [NAD(P)+] | P58143 | 36.7 | 5.65 | 12 | 45.9 | + | |||||||||
67 | Tagatose 1,6-diphosphate aldolase 2 | Q8K5U9 | 36.5 | 4.93 | 15 | 50.5 | + | |||||||||
Arginine degradation | 12 | Ornithine carbamoyltransferase, catabolic | Q8P052 | 37.8 | 5.19 | 19 | 62.8 | + | + | + | + | + | + | + | + | |
40f | Putative carbamate kinase | Q8K6Q9 | 33.2 | 4.71 | 12 | 44.3 | + | + | + | + | + | + | ||||
Amino acid biosynthesis | 14 | Putative branched-chain-amino-acid aminotransferase | Q8K7U5 | 37.2 | 4.90 | 6 | 21.4 | + | + | |||||||
72 | Putative glutamine synthetase | Q8NZG4 | 50.5 | 5.21 | 8 | 19.6 | + | + | ||||||||
Fatty acid and phospholipid biosynthesis | 47 | Putative malonyl coenzyme A-acyl carrier protein transacylase | Q879J3 | 34.5 | 6.02 | 5 | 19.1 | + | + | |||||||
Pantothenate (vitamin B5) biosynthesis | 19 | Putative 2-dehydropantoate 2-reductase (ketopantoate reductase) | Q8P1F1 | 33.8 | 4.93 | 6 | 24.8 | + | + | + | ||||||
Pyridoxine (vitamin B6) biosynthesis | 36 | Putative pyridoxamine-phosphate oxidase | Q8K7X7 | 14.9 | 5.87 | 10 | 68.4 | + | ||||||||
Nucleoside metabolism | 26 | Putative purine nucleoside phosphorylase | Q878J4 | 28.5 | 4.98 | 12 | 44.4 | + | + | + | + | + | + | |||
34 | Uracil phosphoribosyltrans- ferase (UMP pyrophosphorylase) | Q9A194 | 22.8 | 6.30 | 13 | 57.4 | + | + | ||||||||
55 | GMP synthase (gluta- mine hydrolyzing) | Q8K7E6 | 57.5 | 4.91 | 7 | 14.8 | + | |||||||||
63 | Adenylosuccinate synthetase | Q8P2U1 | 47.4 | 5.29 | 10 | 30.7 | + | |||||||||
Metabolic enzyme | 16 | Putative phosphotransacetylase | Q878S0 | 35.9 | 5.08 | 13 | 43.8 | + | + | + | + | |||||
66 | Putative acetoin reduc- tase | Q8P1U1 | 26.8 | 4.79 | 9 | 44.5 | + | |||||||||
68 | Probable manganese-dependent inorganic pyrophosphatase | Q9A1A2 | 33.6 | 4.47 | 7 | 23.8 | + | |||||||||
Virulence factor | 8 | Enolase (2-phosphoglycerate dehydratase) | P82479 | 47.2 | 4.74 | 28 | 65.4 | + | + | + | + | + | + | + | + | + |
9 | Arginine deiminasei | Q8K5F0 | 46.1 | 4.99 | 31 | 70.5 | + | + | + | + | + | + | + | + | ||
13 | GAPDH | P50467 | 35.8 | 5.34 | 18 | 61.8 | + | + | + | + | + | + | + | + | ||
18f | Cysteine protease SpeB precursor | Q93LQ2 | 37.3 | 7.21 | 7 | 22.4 | + | + | ||||||||
23 | Putative C3-degrading proteinase | Q99Y63 | 28.6 | 4.89 | 10 | 43.2 | + | |||||||||
31 | Superoxide dismutase (Mn) | Q8P0D4 | 22.5 | 4.87 | 13 | 81.5 | + | + | + | + | + | |||||
43f | Pyrogenic exotoxin M (SpeM) (fragment) | Q7WYA3 | 24.1 | 7.87 | 6 | 25.0 | + | |||||||||
48fg | FcrA protein precursor | Q54859 | 45.4 | 6.47 | 6 | 15.4 | + | + | ||||||||
50f,g | M protein | Q54840 | 61.7 | 6.24 | 4 | 6.5 | + | + | ||||||||
73 | M protein (fragment) | Q93LJ0 | 27.2 | 5.22 | 6 | 25.0 | + | |||||||||
74 | M protein (fragment) | O86065 | 21.0 | 5.41 | 5 | 21.7 | + | |||||||||
Protein biosynthesis | 7 | Elongation factor Tu | Q8K872 | 43.8 | 4.91 | 24 | 58.3 | + | + | + | + | + | ||||
29 | Peptide deformylase | Q8NZB7 | 22.9 | 5.51 | 11 | 72.5 | + | + | ||||||||
33 | Ribosome recycling factor | Q8P274 | 20.5 | 5.68 | 11 | 64.3 | + | + | + | + | ||||||
38 | Elongation factor G | P82477 | 76.4 | 4.83 | 13 | 22.9 | + | + | ||||||||
44 | Elongation factor Ts | Q8K5L1 | 37.3 | 4.86 | 13 | 45.7 | + | + | + | + | ||||||
57 | Probable sigma54 modulation protein (fragments) | P82482 | 18.4 | 4.45 | 4 | 30.6 | + | |||||||||
62 | Seryl-tRNA synthetase | Q8K635 | 48.1 | 5.17 | 13 | 38.8 | + | |||||||||
64 | Elongation factor P | P82459 | 20.5 | 4.85 | 4 | 29.2 | + | |||||||||
Protein transport | 22f | Putative ABC trans- porter, substrate- binding protein | Q8P2K8 | 30.6 | 7.69 | 7 | 28.6 | + | + | |||||||
39 | Trigger factor | Q879L7 | 47.1 | 4.39 | 11 | 27.6 | + | + | + | |||||||
53 | Putative ABC trans- porter, ATP-binding protein | Q99XH2 | 60.7 | 4.77 | 9 | 21.2 | + | + | ||||||||
65 | Putative copper homeostasis protein (hypothetical protein) | Q8K8H0 | 22.6 | 4.79 | 4 | 24.4 | + | |||||||||
Proteolysis and peptidolysis | 6 | Putative dipeptidase | Q8K7L6 | 51.4 | 4.81 | 12 | 33.0 | + | + | + | + | + | + | + | + | |
10 | Putative X-His dipeptidase | Q99YT8 | 49.1 | 4.74 | 21 | 46.5 | + | |||||||||
30 | Pyrrolidone-carboxylate peptidase | Q8K8C4 | 23.2 | 5.64 | 7 | 38.6 | + | + | ||||||||
45 | Putative methionine aminopeptidase | Q8K718 | 31.6 | 4.84 | 5 | 31.5 | + | |||||||||
Chaperone | 2 | Chaperone protein DnaK | P95831 | 64.8 | 4.62 | 24 | 47.6 | + | + | + | + | + | + | + | + | |
3 | 60-kDa chaperonin GroEL | Q8K5M5 | 56.9 | 4.75 | 35 | 69.6 | + | + | + | + | + | + | + | |||
Stress protein | 35 | Putative alkyl hydroperoxidase | Q99XR7 | 20.5 | 4.65 | 9 | 58.6 | + | + | + | + | + | ||||
49 | Putative glutathione reductase | Q8P1H3 | 48.9 | 5.66 | 5 | 11.1 | + | + | ||||||||
Transcription factor | 58 | Transcription elongation factor GreA | Q9A1C4 | 17.7 | 4.67 | 7 | 61.3 | + | ||||||||
Adhesin | 59f,g | Collagen-like protein B (fragment) | Q9AGC4 | 46.7 | 4.60 | 4 | 10.8 | + | ||||||||
Housekeeping | 15 | Putative alcohol dehydrogenase I | Q9A1X7 | 35.4 | 4.89 | 6 | 20.7 | + | ||||||||
24 | Putative phosphoprotein phosphatase | Q99YM9 | 27.0 | 4.60 | 19 | 80.9 | + | + | ||||||||
25 | Adenylate kinase | Q8K8X1 | 23.7 | 4.83 | 13 | 79.7 | + | + | ||||||||
Unknown function | 20 | Hypothetical UPF0082 protein SPy0316/SpyM3_0231/SPs16 28/spyM18_0311 | Q9A1E6 | 25.9 | 4.49 | 6 | 29.0 | + | ||||||||
37 | Hypothetical phage protein spyM18_0356 | Q8P2H6 | 25.1 | 5.37 | 4 | 13.6 | + | |||||||||
42 | Hypothetical phage protein spyM18_1764 | Q8NZS3 | 22.7 | 4.63 | 3 | 20.2 | + | |||||||||
46 | Hypothetical protein SPy1262 | Q99ZE5 | 19.9 | 4.93 | 9 | 42.5 | + | + | + | |||||||
51 | Hypothetical protein SpyM3_0548 | Q8K7Z2 | 15.4 | 6.05 | 4 | 21.2 | + | |||||||||
56 | Conserved hypothetical protein SPs1095 | Q878P1 | 17.5 | 4.65 | 6 | 54.6 | + | |||||||||
70 | Conserved protein SpyM18_1567 | Q8P050 | 26.6 | 4.67 | 5 | 28.6 | + | |||||||||
71 | Hypothetical protein (phage associated) SPs0647 | Q879B2 | 26.8 | 5.10 | 5 | 21.0 | + |
Identified proteins are indicated by a plus sign.
Swiss-Prot or TrEMBL accession number.
Theoretical values obtained from Swiss-Prot or TrEMBL database.
Number of tryptic peptides detected by MALDI-TOF MS that could be matched to the protein.
Percentage of the protein sequence covered by the matched peptides.
Contains a putative secretion signal sequence identified by SignalP3.0 signal peptide prediction server (http://www.cbs.dtu.dk/services/SignalP/).
Contains a C-terminal LPXTG membrane anchor motif identified by Pfam motif search (http://pfam.wustl.edu/hmmsearch.shtml).