A Multiplexed Parallel Reaction Monitoring Assay to monitor bovine Pregnancy-Associated Glycoproteins throughout pregnancy and after gestation

ABSTRACT: Bovine pregnancy-associated glycoproteins (boPAGs) are extensively glycosylated secretory proteins of trophoblast cells. Roughly 20 different boPAG members are known but their distribution patterns and degree of glycosylation during pregnancy are not well characterized. The objective of the present study was the development of a parallel reaction monitoring-based assay for the profiling of different boPAGs during pregnancy and after gestation. Furthermore, we investigated the effects of N-glycosylation on our analytical results. BoPAGs were purified from cotyledons of four different pregnancy stages. The assay detects 25 proteotypic peptides from 18 boPAGs in a single run. The significantly highest protein abundances were found for boPAG 1 in both, glycosylated and deglycosylated samples. Strongest effects of glycosylation were detected during mid and late pregnancy as well as in afterbirth samples. Furthermore, we identified different boPAG-clusters based on the observed relative protein abundances between glycosylated and deglycosylated samples. A linkage between the impact of glycosylation and potential N-glycosylation sites or phylogenetic relation was not detected. In conclusion, the results of this study provide new starting points to address further research on boPAGs to better understand their physiological role during pregnancy.

INSTRUMENT(S): TripleTOF 5600, Q Exactive

ORGANISM(S): Bos Taurus (bovine) Drosophila Melanogaster (fruit Fly)

TISSUE(S): Placenta

SUBMITTER: Christof Lenz  

LAB HEAD: Christof Lenz

PROVIDER: PXD027383 | Pride | 2022-07-05

REPOSITORIES: Pride