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. 2018 Nov 16;8:407. doi: 10.3389/fcimb.2018.00407

Table 3.

Summary of mass spectrometric results and corresponding identifications of protein spots with significant (P < 0.05) upregulation in fluorescent gel images of the virulent (25) and attenuated (303) Histomonas meleagridis strain.

Spot IDa Protein identity- Species-Contig IDb; Accession Nr. Identification method Gel imagec Theor. pI/ Mrd 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.

a

Spot ID = a unique number assigned to each protein spot by Delta2D software version 4.7 (Decodon GmbH, Greifswald, Germany).

b

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).

c

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.

d

The theoretical isoelectric point (pI) and molecular mass (Mr) of each protein.

e

The fold upregulation of each protein spot retrieved from Delta2D software version 4.7 (Decodon GmbH).

f

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).

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