@article { , title = {N-linked glycosylation regulates human proteinase-activated receptor-1 cell surface expression and disarming via neutrophil proteinases and thermolysin}, abstract = {Proteinase-activated receptor 1 (PAR1) induces activation of platelet and vascular cells after proteolytic cleavage of its extracellular N terminus by thrombin. In pathological situations, other proteinases may be generated in the circulation and might modify the responses of PAR1 by cleaving extracellular domains. In this study, epitope-tagged wild-type human PAR1 (hPAR1) and a panel of N-linked glycosylation-deficient mutant receptors were permanently expressed in epithelial cells (Kirsten murine sarcoma virus-transformed rat kidney cells and CHO cells). We have analyzed the role of N-linked glycosylation in regulating proteinase activation/disarming and cell global expression of hPAR1. We reported for the first time that glycosylation in the N terminus of hPAR1 downstream of the tethered ligand (especially Asn75) governs receptor disarming to trypsin, thermolysin, and the neutrophil proteinases elastase and proteinase 3 but not cathepsin G. In addition, hPAR1 is heavily N-linked glycosylated and sialylated in epithelial cell lines, and glycosylation occurs at all five consensus sites, namely, Asn35, Asn62, Asn75, Asn250, and Asn259. Removing these N-linked glycosylation sequons affected hPAR1 cell surface expression to varying degrees, and N-linked glycosylation at extracellular loop 2 (especially Asn250) of hPAR1 is essential for optimal receptor cell surface expression and receptor stability.}, doi = {10.1074/jbc.M110.204271}, eissn = {1083-351X}, issn = {0021-9258}, issue = {26}, journal = {Journal of biological chemistry}, pages = {22991-23002}, publicationstatus = {Published}, publisher = {American Society for Biochemistry and Molecular Biology}, url = {https://hull-repository.worktribe.com/output/473014}, volume = {286}, keyword = {Health and Health Inequalities, Cell Biology, Biochemistry, Molecular Biology}, year = {2011}, author = {Xiao, Yu Pei and Morice, Alyn H. and Compton, Steven J. and Sadofsky, Laura} }