Table 1. Qualitative analysis of phosphoproteins with phosphorylation site detected.
Phosphorylation sites within peptide sequences and number of samples in which corresponding phosphoprotein was detected after analysis of the fraction bound to TiO2 in healthy (H) and babesiosis-affected dogs (B). #Unique and #PSM correspond to number of unique peptides and spectral counts for a comparison of 10 healthy samples vs. 10 babesiosis samples respectively. NetPhos 3.1 scores and corresponding predicted kinases are shown for each phosphorylation site. Kinases: CaMKII: Ca2+/calmodulin-dependent protein kinase II; CKI: Protein kinase CK1; CKII, Protein kinase CKII; DNAPK: DNA-dependent protein kinase; EGFR: Epidermal growth factor receptor; GSK3: Glycogen synthase kinase 3; INSR: Insulin receptor; p38MAPK: p38 mitogen-activated protein kinase, PKA: Protein kinase A; PKC: Protein kinase C; PKG: Protein kinase G; RSK, ribosomal s6 kinase. P, detected only in pools.
| Protein | Phosphosite | Peptide | H | B | #Unique peptides | #PSM H-B | Accession | ||
|---|---|---|---|---|---|---|---|---|---|
| α-2- antiplasmin ** | S450 |
kEQQDsPDDRDYFQNr / 0.508 CKII |
5 | 10 | 2–3 | 41–41 | 73967363 |
||
| S480 |
lAPPsEEDYPQLHSPk / 0.498 CKI |
5 | 5 | 2–3 | 41–41 | ||||
| Y484 |
lAPPSEEDyPQLHSPk0/ 0.504 INSR |
5 | 5 | 2–3 | 41–41 | ||||
| α-2-HS-Glycoprotein** | S138 |
cDSSPDsAEDVRk / 0.526 CKI |
10 | 10 | 11–14 | 2664–1570 | 545553759 | ||
| S323 |
hAFMGVAsVESASGEAFHVGk / 0.458 CaMK-II |
10 | 10 | 11–14 | 2664–1570 | ||||
| S326 |
hAFMGVASVEsASGEAFHVGk/ 0.484 CaM-II, 0.440 GSK3 |
10 | 5 | 11–14 | 2664–1570 | ||||
| S328 |
hAFMGVASVESAsGEAFHVGk/ 0.428 GSK3 , |
10 | 0 | 11–0 | 2664–0 | ||||
| Chromogranin A* | S190 |
hPDsQAEEDSEGLSQGLVDTEk/ 0.602 DNAPK, 0.595 CKII |
4 | 1 | 1–1 | 4–1 | 244539517 | ||
| Apolipoprotein A1** | S211 |
eGGGAsLAEYHAr / 0.645 PKA, 0.482 CKII |
10 | 10 | 31–39 | 459–1005 | 928133662 | ||
| S220 |
eGGGASLAEYHARAsEQLSALGEk / 0.619 PKA |
5 | 0 | 31–0 | 459–0 | ||||
| Coagulation factor V** | S931 |
wHLVsEKGSYEIVPDAEDMAVDk / 0.513 PKC |
P | P | 1 | 3 | 545504920 | ||
| S935 |
wHLVSEKGsYEIVPDAEDMAVDk / 0.472 GSK3 , |
P | P | 1 | 3 | ||||
| Y936 |
wHLVSEKGSyEIVPDAEDMAVDk / 0.508 EGFR k |
P | P | 1 | 3 | ||||
| Complement factor H** | S883 |
sSIFsEEIEETSKPk / 0.629 CKII, 0.571 CKI |
10 | 10 | 3–14 | 37–48 | 74005944 | ||
| DNA directed RNA* polymerase subunit RPA43 |
S300 |
kKHQEVQDQDPVFQGSDSsGYQSDHk / 0.857 PKC, 0.524 CKII |
0 | 2 | 0–1 | 0–6 | Q3B726 | ||
| Fibrinopeptide A/Fibrinogen α ** | T20 |
tNSKEGEFIAEGGGVr / 0.491 CKII 0.481 cdc2 |
9 | 3 | 1–1 | 46–11 | 73978329 |
||
| S22 |
tNsKEGEFIAEGGGVr / 0.563 CKI 0.458 CaMKII |
9 | 3 | 2–1 | 46–11 | ||||
| Fibrinopeptide B/Fibrinogen β ** | Y31 |
hyYDDTDEEEr / 0.377 INSR |
4 | 5 | 1–1 | 30–53 | 545524893 |
||
| Y32 |
hYyDDTDEEEr / 0.380 INSR , |
0 | 2 | 1–1 | 30–53 | ||||
| T35 |
hYYDDtDEEEr / 0.734 CKII |
4 | 5 | 1–1 | 30–53 | ||||
| Fibrinogen γ ** | S128 |
yEALVGsHESNIr / 0.477 cdc2 |
P | P | 2 | 4 | 73977992 | ||
| Fibronectin** | S2384 |
rTNTNVNCPIECFMPLDVQADREDsRe 0.559 RSK |
10 | 10 | 8–5 | 58–42 |
P02751 |
||
| Immunoglobulin heavy chain variable region ** | T47 |
rLSCVASGFtLSDYGLSWVr/ 0.476 CaM-II |
P | P | 1 | 3 | 208342066 | ||
| Insulin-like growth factor binding protein 3 * | S201 |
vDYESQSTDTQNFsSEYk/ 0.521 CKII, 0.519 CKI |
3 | 0 | 1–0 | 6–0 | 359321488 | ||
| Inter-alpha-trypsin inhibitor heavy chain H2 (ITIH2)** | S60 |
sVFGEsGEVMEEADQVTLYSYk / 0.589 CKII |
P | P | 11 | 17 |
73949158 | ||
| Histidine-rich glycoprotein** |
S464 |
qGQGPPPQHsEEr / 0.448 GSK3 |
5 | 5 | 3–3 | 70–42 | 545553762 | ||
| Kanadaptin* | T625 |
lQQEtELEEAVQDTRPPTDLMCSKETk / 0.714 CKII |
2 | 0 | 1–0 | 4–0 | 545527407 |
||
| T634 |
lQQETELEEAVQDtRPPTDLMCSKETk / 0.495 cdc2 |
2 | 0 | 1–0 | 4–0 | ||||
| Kininogen 1 X1** | T326 |
etMCSKESNEELAESCQINk / 0.600 PKC |
10 | 10 | 7–12 | 48–38 | 345796419 | ||
| S332 |
eTMCSKEsNEELAESCQINk / 0.660 CKII , |
10 | 0 | 9–0 | 230–0 | ||||
| X2** | T326 |
etMCSKESNEELAESCQINk / 0.600 PKC |
10 | 10 | 3–10 | 35–147 | 57109938 | ||
| S332 |
esNEELAESCQINk / 0.590 SRC |
10 | 0 | 6–0 | 164–0 | ||||
| Kinesin-like KIF26A* |
S1726 |
lGRKPsLPGQWVDLPPPLAGSLk / 0.823 PKA, 0.542 PKG , |
0 | 2 | 1–1 | 1–2 | Q9ULI4 | ||
| Serum albumin** | T267 |
vHtECCHGDLLECADDRADLAk / 0.451 CKII |
10 | 10 | 3–4 | 50–73 | P02768-1 | ||
| Serum albumin X1** | S494 | mSCADDFLSVVLNRLCVLHEKTPVsEr/0.743 PKC | P | P | 2 | 3 | 545520919 | ||
| Serum amyloid A** | S101 |
fGDSGHGAEDsKADQAANEWGr / 0.461 cdc2 EWGRSGKDPN HFRPAGLPDK Y |
0 | 10 | 0–8 | 0–382 | 545536980 |
||
| Sulfhydryl oxidase** | S398 | hNLDHsQETAEAQEVLQAIr / 0.649 DNAPK | 6 | 9 | 1–1 | 9–24 | 928139154 |
||
| T401 | hNLDHSQEtAEAQEVLQAIr / 0.649 DNAPK | 6 | 9 | 1–1 | 9–24 | ||||
| Golgi membrane protein 1** | S276 |
gETNEIQVTsEEEPQr / 0.658 CKII , |
P | P | 1 | 2 | 928125053 | ||
| Vitronectin** | Y75 |
rGDVFTMPEDEyTVYDDGEEk / 0.634 SRC |
P | P | 6 | 9 | 73966959 | ||
*, False Discovery Rate 5%
**, False Discovery Rate 1%