Proteomic analysis of bovine conceptus fluids during early pregnancy.
A proteomic analysis of bovine amniotic and allantoic fluids collected around Day 45 of gestation was performed using gel-based and LC-based MS workflows. A depletion/enrichment protocol using ultrafiltration under denaturing and reducing conditions produced an enriched fraction containing protein species predominantly between 5 and 50 kDa molecular weight. The analyses of conceptus fluid proteins were performed using two strategies; first, 2-DE coupled with MALDI-TOF-MS/MS and LC-ESI-MS/MS analysis of individual protein spots and second, a global protein snapshot of the enriched 5-50 kDa protein fraction by LC-ESI-MS/MS and LC-MALDI-TOF-MS/MS. Allocation of bovine specific protein identities was achieved by searching the Interactive Bovine In Silico SNP (IBISS) and NCBInr protein sequence databases resulting in the confident PMF identification and MS/MS confirmation of >200 2-DE generated allantoic fluids protein spots (74 individual protein species identified) and the MS/MS peptide identification of 105 LC-ESI-MS/MS generated protein identities. In total, the identity of 139 individual protein species from allantoic fluids was confirmed with peptide sequence probability MOWSE scores at the p<0.05 level or better. The comparison of bovine Day 45 amniotic and allantoic fluids protein profiles revealed differences between these two conceptus fluids in early pregnancy.
Riding
George A
GA
CSIRO Livestock Industries, Queensland Bioscience Precinct, St Lucia, QLD 4067, Australia. george.riding@csiro.au
Jones
Alun
A
Holland
Michael K
MK
Hill
Jonathan R
JR
Lehnert
Sigrid A
SA
D000482:Allantois / Q000737:chemistry* / Q000378:metabolism; D000653:Amniotic Fluid / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D008431:Maternal-Fetal Exchange / Q000502:physiology; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000737:chemistry* / Q000378:metabolism; D040081:Protein Array Analysis; D040901:Proteomics; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011257:Pregnancy Proteins