Table 3.
Spot IDa | Protein identity- Species-Contig IDb; Accession Nr. | Identification method | Gel imagec | Theor. pI/ M | Fold upregulatione | Protein scoreF | Nr. of peptides |
---|---|---|---|---|---|---|---|
CARBOHYDRATE METABOLISM/ PLASMINOGEN (PLG)-BINDING: VIRULENT H. meleagridis | |||||||
610 | Fructose-bisphosphate aldolase (FBAL)- Contig601; HAGI01000595 | MALDI-TOF/TOF | 25 | 6.0/37.2 | 2.726 | 385 | 6 |
526 | Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)- Contig449; HAGI01000443 | MALDI-TOF/TOF | 25 | 6.6/40.0 | 2.708 | 371 | 4 |
509 | Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)- Contig449; HAGI01000443 | MALDI-TOF/TOF | 25 | 6.6/40.0 | 1.856 | 445 | 4 |
524 | Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)- Contig449; HAGI01000443 | MALDI-TOF/TOF | 25 | 6.6/40.0 | 1.811 | 318 | 4 |
CYTOSKELETON/PLASMINOGEN (PLG)-BINDING: VIRULENT H. meleagridis | |||||||
535 | Actin- Contig2112; HAGI01002078 | MALDI-TOF/TOF | 25 | 5.1/42.5 | 2.942 | 124 | 2 |
627 | Actin- Contig2112; HAGI01002078 | MALDI-TOF/TOF | 25 | 5.1/42.5 | 2.725 | 169 | 2 |
386163 | Actin family protein- Contig444; HAGI01000438 | MALDI-TOF/TOF | 25 | 6.5/44.6 | 2.687 | 128 | 2 |
CARBOHYDRATE METABOLISM: VIRULENT H. meleagridis | |||||||
202345 | Phosphoenolpyruvate carboxykinase (PEPCK)- Contig1899; HAGI01001872 | MALDI-TOF/TOF | 25 | 5.8/67.1 | 2.400 | 415 | 5 |
63267 | Iron-containing alcohol dehydrogenase (ADH)- Contig362; HAGI01000356 | LC-MS/MS | 25 | 5.6/43.5 | 2.259 | 246 | 17 |
640 | Phosphoglycerate mutase (PGlyM)- Contig1739; HAGI01001716 | MALDI-TOF/TOF | 25 | 6.8/29.8 | 1.602 | 294 | 4 |
METABOLIC PROCESSES: VIRULENT H. meleagridis | |||||||
42635 | Clan MG, family M24, aminopeptidase P-like metallopeptidase- Contig29; HAGI01000029 | MALDI-TOF/TOF | 25 | 5.2/51.9 | 2.510 | 101 | 1 |
356639 | Clan MG, family M24, aminopeptidase P-like metallopeptidase- Contig29; HAGI01000029 | LC-MS/MS | 25 | 5.2/51.9 | 1.345 | 102 | 8 |
ADAPTATION TO STRESS: BACTERIAL | |||||||
417 | 60 kDa chaperonin- Escherichia coli- Q6Q099_ECOLX | LC-MS/MS | 25 | 4.8/57.3 | 1.535 | 305 | 21 |
CARBOHYDRATE METABOLISM: BACTERIAL | |||||||
197187 | Class II fructose-bisphosphate aldolase- E. coli- A0A0D8VY27_ECOLX | MALDI-TOF/TOF | 25 | 5.5/39.1 | 1.404 | 220 | 2 |
CYTOSKELETON ORGANIZATION: ATTENUATED H. meleagridis | |||||||
408 | Coronin- Contig2106; HAGI01002072 | MALDI-TOF/TOF | 303 | 5.9/50.2 | 1.492 | 525 | 5 |
409 | Coronin- Contig1193; HAGI01001177 | MALDI-TOF/TOF | 303 | 5.9/50.3 | 1.395 | 716 | 7 |
METABOLIC PROCESSES: ATTENUATED H. meleagridis | |||||||
587944-1 | Transketolase family protein- Contig1077; HAGI01001062 | MALDI-TOF/TOF | 303 | 5.4/62.7 | 1.551 | 231 | 3 |
587944-2 | Transketolase family protein- Contig1077; HAGI01001062 | MALDI-TOF/TOF | 303-Gel4 | 5.4/62.7 | 1.551 | 246 | 4 |
CARBOHYDRATE METABOLISM/PLASMINOGEN (PLG)-BINDING: ATTENUATED H. meleagridis | |||||||
495 | Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)- Contig449; HAGI01000443 | MALDI-TOF/TOF | 303 | 6.6/40.0 | 1.460 | 455 | 4 |
METABOLIC PROCESSES: BACTERIAL | |||||||
153815 | Bifunctional purine biosynthesis protein purH- E. coli- A0A085PAJ0_ECOLX | MALDI-TOF/TOF | 303 | 5.5/57.3 | 1.414 | 502 | 6 |
Results were obtained by the two-dimensional differential gel electrophoresis (2D-DIGE) experiment, designated as 2D-DIGE-β. The identified proteins were categorized according to their proposed function and listed within categories according to their fold upregulation (from high to low).
Spots of interest were excised and pooled from all four silver-stained fluorescent gels included in the 2D-DIGE-β experiment.
Proteins that take part in metabolic processes other than carbohydrate metabolism are grouped together.
Spot ID = a unique number assigned to each protein spot by Delta2D software version 4.7 (Decodon GmbH, Greifswald, Germany).
Contig identification number (ID) was obtained from the de novo transcriptome sequencing of a virulent and an attenuated H. meleagridis strain (Mazumdar et al., 2017).
Gel image = 25: the protein spot was significantly upregulated in fluorescent gel images displaying proteins of the cultivated virulent strain.
303: the protein spot was significantly upregulated in fluorescent gel images displaying proteins of the cultivated attenuated strain.
587944-2: The spot was excised and analyzed separately from Gel 4.
The theoretical isoelectric point (pI) and molecular mass (Mr) of each protein.
The fold upregulation of each protein spot retrieved from Delta2D software version 4.7 (Decodon GmbH).
Protein score = MALDI-TOF/TOF identifications with protein score >80 are significant in MASCOT and fulfill the stricter criteria of ProteinScape 2.1 software.
The following protein spots were not identified: 457 (gel image: 25, fold upregulation: 1.255) and 63593 (gel image: 25, fold upregulation: 3.443).