Jin Young Gil, Jeong-Nam Park, Kyung Jin Lee, Ohsuk Kwon, Hye-Shin Chung, Hyun Ah Kang, Doo-Byoung Oh
언어
영어(ENG)
URL
https://www.earticle.net/Article/A212723
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원문정보
초록
영어
Some portions of N-glycans are phosphorylated in Saccharomyces cerevisiae by the dual action of two proteins encoded by MNN4 (ScMNN4) and MNN6 (ScMNN6). It has been known that ScMnn4 protein (ScMnn4p) is a positive regulator of ScMnn6p, a real mannosyltransferase adding mannosylphosphate residues to glycan. On the other hand, YlMPO1, Yarrowia lypolytica homologue of ScMNN4, was reported to mediate mannosyl-phosphorylation of N- and O-glycans in Y. lypolytica without the involvement of MNN6 homologues (1). In the present study, we further tested whether YlMpo1p can display mannosyltransferase activity independently in S. cerevisiae without the help of either ScMnn4p or ScMnn6p. YlMPO1 gene was heterologously expressed in S. cerevisiae wild-type, ScMNN4 disruptant (ScMnn4Δ), ScMNN6 disruptant (ScMnn6Δ), ScMNN4 and ScMNN6 double disruptant (ScMnn4ΔMnn6Δ). N-glycan profiles of their cell wall mannoproteins, which were analyzed using HPLC and DNA sequencer, clearly showed that the expression of YlMPO1 greatly increases the amount of mannosyl-phosphorylated glycans in all these mutant strains as well as wild-type, indicating its independent mannosyltransferase activity. Furthermore, increased amounts of mannosyl-phosphorylated glycans were much higher upon the heterologous expression of YlMPO1 compared to recombinant expression of ScMNN4. The strategy of YlMPO1 expression to increase the phosphorylated glycans would be usefully employed for glyco-engineering of yeast to produce therapeutic enzymes containing mannose-6-phosphate.
저자
Jin Young Gil [ Korea Research Institute of Bioscience & Biotechnology, Department of Biotechnology, Hannam University ]
Jeong-Nam Park [ Department of Life Science, Chung-Ang University, Seoul 156-756, Korea ]
Kyung Jin Lee [ Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea ]
Ohsuk Kwon [ Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea ]
Hye-Shin Chung [ Department of Biotechnology, Hannam University, Daejeon 305-811, Korea ]
Hyun Ah Kang [ Department of Life Science, Chung-Ang University, Seoul 156-756, Korea ]
Doo-Byoung Oh [ Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, 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.