Table 3.
Comparison between relevant PPRSV-related and pig proteins identified in other studies and the discriminatory peaks found in this study
| Method of identification of the peak [reference] | Protein | Reported MW (kDa) | Regulation in other studies | MW (kDa) of the peak identified in this study with a difference ≤0.3 kDa compared to the other reports (regulation PRRSV-positive vs. -negative) |
|---|---|---|---|---|
|
PRRSV proteins |
|
|
|
|
| - Calculated molecular mass from amino acid sequence [23,24] |
ORF1a – non structural polyprotein |
260 - 270 |
|
|
| - Calculated molecular mass from amino acid sequence [23,24] |
ORF1ab – non structural polyprotein |
420 - 430 |
|
|
| - Estimated size from amino acid sequence [25] |
ORF2a - glycoprotein 2a (GP2a) |
28.4 |
|
|
| - 2-DE PAGE and MALDI-TOF [26] |
|
29.4 |
|
|
| - SDS page and western of MARC-145 cells infected with PRRSV [27] |
ORF2b - non-glycosylated protein 2b |
10 |
|
10.041 (up-regulated) |
| - Estimated size from amino acid sequence [25] |
ORF3 - glycoprotein 3 (GP3) |
30.6 |
|
|
| - 2-DE PAGE and MALDI-TOF [26] |
|
29 |
|
|
| - Estimated size from amino acid sequence [25] |
ORF4 - glycoprotein 4 (GP4) |
20 |
|
19.761 (up-regulated) |
| - 2-DE PAGE and MALDI-TOF [26] |
|
19.5 |
|
19.761 (up-regulated) |
| - Estimated size from amino acid sequence [25] |
ORF5 - glycoprotein 5 (GP5, E) |
22.4 |
|
|
| - 2-DE PAGE and MALDI-TOF [26] |
|
22.4 |
|
|
| - Estimated size from amino acid sequence [25] |
ORF6 - matrix protein (M) |
18.9 |
|
|
| - 2-DE PAGE and MALDI-TOF [26] |
|
19 |
|
|
| - Estimated size from amino acid sequence [25] |
ORF7 - nucleocapsid protein (N) |
13.8 |
|
13.785 (up-regulated) |
| - 2-DE PAGE and MALDI-TOF [26] |
|
13.5 |
|
13.785 (up-regulated) |
|
Pig protein peaks related to PRRSV infection |
|
|
|
|
| - MALDI-TOF (sera of pigs after few days of infection with PRRSV vs. normal) [28] |
Alpha 1 S (a1S)-subunit of porcine Haptoglobin (Hp) |
9.244 |
Up-regulated in PRRSV infected sera (after 1–7 days) |
9.136 (up-regulated) |
| |
Unknown peak |
4.165 |
No difference |
4.161 (up-regulated) |
| |
Unknown peak |
4.460 |
No difference |
4.458; 4.462 (both up-regulated) |
| |
Unknown peak |
5.560 |
No difference |
5.536 (down-regulated) |
| |
Unknown peak |
8.330 |
No difference |
8.328 (up-regulated) |
| |
Unknown peak |
8.825 |
No difference |
8.843 (up-regulated) |
| |
Unknown peak |
12.250/12.55 |
No difference |
12.237/12.522 (both up-regulated) |
| |
Unknown peak |
14.010 |
No difference |
13.785 (up-regulated) |
| - 2-DE PAGE and MALDI-TOF of cellular proteins incorporated in PRRSV virions [26] |
Keratin 10 |
58.8 |
|
|
| |
Coronin, actin binding protein, 1B |
55.7 |
|
|
| |
Keratin 9 |
62 |
|
|
| |
Tubulin, beta polypeptide |
47.7 |
|
|
| |
Tubulin, alpha, ubiquitous |
50.1 |
|
|
| |
Beta-actin |
41.7 |
|
|
| |
Actin, gamma 1 propeptide |
41.8 |
|
|
| |
Keratin 1 |
66 |
|
|
| |
Tropomyosin 1 alpha chain isoform 4 |
32.9 |
|
|
| |
Cofilin 1 (non-muscle) |
18.5 |
|
|
| |
Heat shock 70 kDa protein 8 isoform 1 |
70.8 |
|
|
| |
Heat shock 60 kDa protein 1 |
61 |
|
|
| |
Ribosomal protein P0 |
34.2 |
|
|
| |
Heat shock protein 27 |
22.3 |
|
|
| |
Transketolase |
67.8 |
|
|
| |
Pyruvate kinase |
57.8 |
|
|
| |
Phosphoglycerate dehydrogenase |
56.6 |
|
|
| |
Aldehyde dehydrogenase 1A1 |
54.8 |
|
|
| |
UDP-glucose dehydrogenase |
55 |
|
|
| |
Enolase 1 |
47.1 |
|
|
| |
Phosphoglycerate kinase 1A isoform 2 |
44.6 |
|
|
| |
Glyceraldehyde-3-phosphate dehydrogenase |
23.8 |
|
23.496 (up-regulated) |
| |
Guanine nucleotide binding protein (G protein), beta polypeptide 1 |
37.3 |
|
|
| |
L-lactate dehydrogenase B |
36.6 |
|
|
| |
Chain A, Fidarestat Bound To Human Aldose Reductase |
35.7 |
|
|
| |
PREDICTED: lactate dehydrogenase |
36.6 |
|
|
| |
Peroxiredoxin 1 |
22.1 |
|
|
| |
Proteasome activator hPA28 subunit beta |
27.3 |
|
27.606 (down-regulated) |
| |
Triosephosphate isomerase 1 |
26.6 |
|
|
| |
Chaperonin containing TCP1, subunit 3 (gamma) |
60.4 |
|
|
| |
Chaperonin containing TCP1, subunit 6A (zeta 1) |
58 |
|
|
| |
Chaperonin containing TCP1, subunit 5 (epsilon) protein |
59.6 |
|
|
| |
Chaperonin containing TCP1, subunit 2 |
57.4 |
|
|
| |
PRP19/PSO4 pre-mRNA processing factor 19 homolog |
55.1 |
|
|
| |
Retinoblastoma binding protein 4 isoform a |
47.6 |
|
|
| |
Eukaryotic translation initiation factor 4A isoform 1 |
46.1 |
|
|
| |
Proliferating cell nuclear antigen |
28.7 |
|
|
| |
Alpha2-HS glycoprotein |
35.6 |
|
|
| |
Annexin A2 |
38.5 |
|
|
| |
Annexin A5 |
35.9 |
|
|
| |
Annexin A4 |
36.1 |
|
|
| |
S100 calcium binding protein A10 |
11.2 |
|
11.090 (up-regulated) |
| |
Galectin-1 |
14.7 |
|
14.843 (down-regulated) |
| |
T-complex protein 1 isoform a |
60.3 |
|
|
| |
Gastric-associated differentially expressed protein YA61P |
14.9 |
|
14.843 (down-regulated) |
| - 2-DE PAGE and MALDI-TOF of PAM infected with PRRSV vs. normal [29] |
Lymphocyte cytosolic protein 1 |
70 |
Up-regulated in infected PAM |
|
| |
65 kDa macrophage protein |
70.2 |
Up-regulated |
|
| |
L plastin isoform 2 |
41.4 |
Up-regulated |
|
| |
Enolase 1 |
47.1 |
Up-regulated |
|
| |
BUB3 budding uninhibited by benzimidazoles 3 isoform a |
37.1 |
Up-regulated |
|
| |
Heat shock 27 kDa protein 1 |
22.9 |
Up-regulated |
23.162 (up-regulated) |
| |
Proteasome beta 2 subunit |
22.8 |
Up-regulated |
|
| |
Transgelin 2 |
21.1 |
Up-regulated |
|
| |
NADP-dependent isocitrate dehydrogenase |
46.7 |
Up-regulated |
|
| |
Superoxide dismutase 2 |
11.7 |
Up-regulated |
11.613 (up-regulated) |
| |
Lamin C |
65.1 |
Up-regulated |
|
| |
Aconitase |
98.1 |
Up-regulated |
|
| |
Long chain acyl-CoA dehydrogenase |
47.9 |
Up-regulated |
|
| |
Proteasome subunit alpha type 1 |
29.5 |
Up-regulated |
|
| |
70 kDa heat shock cognate protein atpase domain |
41.9 |
Up-regulated |
|
| |
Similar to dihydrolipoamide S-succinyltransferase (E2 component of 2-oxo-glutarate complex) |
48.9 |
Up-regulated |
|
| |
Similar to cleavage stimulation factor, 3 pre-RNA, subunit 1 isoform 3 |
47.3 |
Up-regulated |
|
| |
Beta Actin |
39.2 |
Down-regulated in infected PAM |
|
| |
Beta Actin |
32.1 |
Down-regulated |
|
| |
Myoglobin |
16.9 |
Down-regulated |
17.171 (up-regulated) |
| |
Vacuolar protein sorting 29 |
20.5 |
Down-regulated |
20.322 (up-regulated) |
| |
Transketolase |
67.9 |
Down-regulated |
|
| |
Eukaryotic translation initiation factor 3, subunit 5 |
37 |
Down-regulated |
|
| |
Cathepsin D protein |
42.7 |
Down-regulated |
|
| |
Similar to lymphocyte-specific protein 1 |
40.9 |
Down-regulated |
|
| - 2-DE PAGE and MALDI-TOF of PAM constitutively expressing the PRRSVN protein vs. normal [30] |
Proteasome subunit alpha type 6 |
28.5 |
Up-regulated in PAM expressing PRRSVN |
|
| |
Heat shock protein 27 kDa |
23 |
Up-regulated |
23.162 (up-regulated) |
| |
Annexin 1 |
38.5 |
Up-regulated |
|
| |
Septin 2 |
42.9 |
Up-regulated |
|
| |
Spermidine synthase |
34.4 |
Down-regulated in PAM expressing PRRSVN |
|
| |
Major vault protein |
19.3 |
Down-regulated |
|
| |
Ferritin L subunit |
18.3 |
Down-regulated |
|
| |
Nucleoside diphosphate kinase A |
17.3 |
Down-regulated |
17.218 (up-regulated) |
| |
Chaperonin containing TCP-1 beta subunit |
57.8 |
Down-regulated |
|
| |
Dihydropyrimidinase related protein 2 |
62.7 |
Down-regulated |
|
| |
Translation elongation factor 2 |
47.2 |
Down-regulated |
|
| - 2-DE PAGE and MALDI-TOF of PAM and Marc-145 cells infected with PRRSV [31] |
Cofilin 1 |
25.773 |
Up-regulated in Marc-145 |
|
| |
Actin-related protein |
16.278 |
Up-regulated in PAM |
|
| |
Vimentin |
30.826 |
Up-regulated in PAM |
|
| |
Alpha cardiac actin |
16.758 |
Up-regulated in PAM |
|
| |
Cofilin 1 |
18.507 |
Up-regulated in PAM |
|
| |
Stress 70 protein |
55.119 |
Up-regulated in Marc-145 |
|
| |
Peroxiredoxin 2 |
19.418 |
Up-regulated in Marc-145 |
|
| |
Heat shock 27 kDa protein 1 |
22.927 |
Up-regulated in Marc-145 |
23.162 (up-regulated) |
| |
Peroxiredoxin 6 |
24.995 |
Up-regulated in Marc-145 |
|
| |
Heat shock protein beta 1 (HSPB1) |
22.768 |
Up-regulated in PAM |
|
| |
Ubiquitin |
8.559 |
Up-regulated in Marc-145 |
8.552 (up-regulated) |
| |
Cystatin B (CSTB) |
25.288 |
Up-regulated in PAM |
|
| |
FYVE finger containing phosphoinositide kinase |
232.904 |
Up-regulated in PAM |
|
| |
Pyruvate kinase isozymes M1/M2 (PKM2) |
57.744 |
Up-regulated in PAM |
|
| |
UPF 0681 protein KIAA1033 |
136.330 |
Up-regulated in Marc-145 |
|
| |
Tropomyosin alpha 4 chain (TPM4) |
28.504 |
Up-regulated in PAM |
|
| |
UPF 0568 protein |
28.191 |
Up-regulated in PAM |
|
| |
LIM and SH3 protein 1 |
29.975 |
Down-regulated in Marc-145 |
|
| |
Plectin 1 |
532.578 |
Down-regulated in Marc-145 |
|
| |
Glial fibrillary acidic protein (GFAP) |
46.497 |
Down-regulated in Marc-145 |
|
| |
Plectin 1 |
516.572 |
Down-regulated in PAM |
|
| |
Galectin 1 |
14.736 |
Down-regulated in Marc-145 |
14.843 (down-regulated) |
| |
Galectin 1 |
14.590 |
Down-regulated in PAM |
|
| |
Superoxide dismutase 1 (SOD1) |
15.236 |
Down-regulated in PAM |
|
| |
Prohibitin |
29.757 |
Down-regulated in Marc-145 |
|
| |
Epidermal fatty acid-binding protein 5 (FABP5) |
15.199 |
Down-regulated in PAM |
|
| |
A kinase anchoring protein AKAP350 |
416.855 |
Down-regulated in PAM |
|
| Pyridoxine 5 phosphate oxidase variant | 29.896 | Down-regulated in PAM |
List of relevant PPRSV and pig proteins that have been shown in other studies and might correspond to the significantly expressed peaks found here with SELDI-TOF MS. The method used in other studies to identify the peak with the corresponding reference, the protein names, as well as their MW and regulation are reported. The last column indicates the MWs (with in parenthesis the regulation in PRRSV-positive compared to PRRSV-negative piglets) of discriminatory peaks identified in this study that showed a difference ≤0.3 kDa compared to the other studies