An Insight into Glyco-microheterogeneity of Plasma von Willebrand Factor (vWF) by Mass Spectrometry

Von Willebrand Factor (vWF), a plasma glycoprotein, plays essential roles in primary hemostasis, in cooperation with other coagulation factors. The glycosylation affects many biological phases during the protein life-cycle. In this work, we comprehensively characterized the microheterogeneity at all probable N-glycosites in simultaneous with O-glycosites, beside the N-occupancy estimation.An RP-LC-MS/MS system functionalized with CID and HCD tandem mass techniques was utilized. Our results showed gradual glycosylation occupancy along the protein backbone. A total of 181 glycoforms, 173 N-froms and 8 O-forms, were detected and specified into plasma vWF glycosites. Given subtle characterization of site-specific glycoforms is critical for profound understanding the protein biological functions and activity.