R306S Gain-of-Function Mutation Enhances PCSK9- mediated Lowering of LDL-R Expression

摘要：

OBJECTIVE To investigate the effect of missense mutation in PCSK9 gene R306S on the metabolism of LDL and to analyze its relationship between structure and function. METHODS The wild type PCSK9 gene and the second or the third grade structures of the mutant R306S coding protein were all forecasted with the aid of Expert Protein Analysis System. The PCSK9 gene eukaryocyte expression vector was constructed and the nucleotide sequence was determined; The pathotype recombinant eukaryotic expression plasmid which carrying PCSK9 gene was constructed by directedmutation method; Liposorne was used as intermediate to transfect the recombinant plasmid into BEL-7402 cell, the cells receiving no plasmid transfection served as control during this process; The LDL-R level was detected by Western Blot. RESULTS Through simulating the second and the third grade structures of protein, conformations changes were found in the two major coding protein parts of mutated PCSK9 gene: carboxy-terminal domain and catalyzing subunit site domain, the space between these two domains increased as well. And that was not found in wild type of PCSK9 gene. Tansfecting wild type of PCSK9 plasmid into BEL-7402 cell could lead to the decrease of LDL-R mature protein expression compared with controlled group(P<0.05) and there was a significant decrease when cells transfected with positive control plasmids (P<0.01) . CONCLUSIONS The R306S mutation, gene mutant of PCSK9, could significantly lead to the decrease of LDL-R mature protein expression. Analysis results obtained from computer analogy confirmed that there were changes on the second and the third grade structures of R306S coding protein.

关键词： proprotein convertase subtilisinAexin type 9 gene familial hypercholesterolemia Conformation simulating LDL receptor gain of function

作者: Jie Lin Youyan Guo Zuo Wang Fengru Shi Ya Yang Qiang Yong Xiaodong Pan Zhisheng Jiang Luya Wang

作者单位: Institute of Cardiovascular Diseases, Key Laboratory for Arteriosclerology of Hunan Province, Univer The Key Laboratory of Remodeling-related Cardiovascular Diseases, Beijing An Zhen Institute of Cardiovascular Diseases, Key Laboratory for Arteriosclerology of Hunan Province, Univer

会议类型: 国际会议

会议名称: 2011 4th International Conference on Biomedical Engineering and Informatics(第四届生物医学工程与信息学国际会议 BMEI 2011)

会议地点: 上海

会议语种:英文

页码: 1617-1620

在线出版日期: 2011-10-15（万方平台首次上网日期，不代表论文的发表时间）