Recombinant coagulation factor IX (rFIX) is an extremely heterogeneous glycoprotein having multiple N-and O-glycosylation which influence on pharmacological functions including biological activity and in vivo stability. The study of glycosylation on rFIX has mainly focused on N-glycans due to an inherent complexity of O-glycosylation and the absence of analytical platform. Here, we comprehensively characterized the O-glycosylation on recombinant coagulation factor IX (rFIX) using MS-based glycomic and glycoproteomic approach. Briefly, O-glycans were chemically released using β-elimination while O-glycopeptides were prepared by pronase digestion to determine site-specific O-glycosylation distribution. Successfully we could identify unique O-glycosylation modifications such as O-fucosylation and O-glucosylation as well as a typical O-glycosylation on rFIX using nanoLC/MS and Tandem MS. Particularly, through O-glycopeptide analysis, we demonstrated that O-glucosylation and O-fucosylation were uniformly displayed on O-sites Serine 53 and 61 of EGF domain, respectively. While core 1-type O-glycosylations were overwhelmingly distributed across other O-sites (Threonine 159, 169, 172, and 179) in activation peptide of rFIX. Site-specific profiling including micro-and macro-heterogeneity could provide macroscopic aspect to figure out O-glycosylation pattern decorated on rFIX.
저자
Youngsuk Seo [ Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon 34134, Korea Asia-Pacific Glycomics Reference Site, Daejeon 34134, Korea ]
Myung Jin Oh [ Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon 34134, Korea Asia-Pacific Glycomics Reference Site, Daejeon 34134, Korea ]
Hyun Joo An [ Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon 34134, Korea Asia-Pacific Glycomics Reference Site, Daejeon 34134, Korea ]
본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.