Kafkas Üniversitesi Veteriner Fakültesi Dergisi Articles in Press
Eukaryotic Expression, Co-IP and MS Identify BMPR-1B Protein-protein Interaction Network
Jianlei JIA1,2, Ziqi ZHANG3, Qian CHEN1, Linsheng GUI1,2, Shengzhen HOU2, Haiqin LI2
1State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining, Qinghai, 810016, PR CHINA
2College of Agriculture and Animal Husbandry, Qinghai University, Xining, Qinghai, 810016, PR CHINA
3College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, Gansu, 730070, PR CHINA
DOI : 10.9775/kvfd.2019.22655 Bone Morphogenetic Protein Receptors 1B (BMPR-1B) belongs to the transforming growth factor β super family and plays a critical role in the embryo, postnatal development, as well as maintaining homeostasis in several organs and tissues by regulating cell differentiation, proliferation, survival and motility. Loss-of-function mutations in BMPR-1B gene exon-8 (FecB locus) lead to an increase in ewe ovulation rate. Herein, we construct eukaryotic expression system and prepared monoclonal antibody, then characterized BMPR-1B protein-protein interactions (PPIs). Using Coimmunoprecipitation coupled to mass spectrometry (IP-MS), we identified 23 proteins that specifically interact with BMPR-1B within ewes" ovary extracts. Select PPIs were validated by immunoblot, demonstrating that BMPR-1B associates with a number of other BMPs primarily via signal transduction in ovary. Analysis of the biological processes and pathways associated with BMPR-1Binteracting proteins indicates enrichment for the reproduction process via association with Smads proteins and TGF-beta signaling pathway via BMP2 and BMP4 pathways. Moreover, several other target proteins, including GDF5, GDF9, RhoD and HSP 10, interact with BMPR-1B. Taken together, our results define a PPI network for BMPR-1B, suggest a novel pathway for this protein, and provide new insights into the contributions of BMPR-1B to litter size. Keywords : Sheep, BMPR-1B, Co-immunoprecipitation, Mass spectrometry, Protein-protein interaction