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Proteomic analysis of bovine conceptus fluids during early pregnancy
Author(s) -
Riding George A.,
Jones Alun,
Holland Michael K.,
Hill Jonathan R.,
Lehnert Sigrid A.
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700465
Subject(s) - conceptus , amniotic fluid , peptide mass fingerprinting , in silico , biology , protein sequencing , proteomics , andrology , chemistry , peptide sequence , chromatography , gestation , pregnancy , biochemistry , fetus , genetics , medicine , gene
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.