摘要
目的:探讨人血浆脂蛋白(a)的赖氨酸结合功能的结构基础。方法:利用基因重组质粒在Ecoli菌株中表达野生型和变异型apo(a)KringleⅣ-10的GST融合蛋白,经Glutathione_Agrose亲和层析纯化,通过Lys_Sepharose亲和柱检测它们的赖氨酸结合能力。结果:野生型kringleⅣ-10的融合蛋白具有较强的赖氨酸结合能力,而变异型kringleⅣ-10的融合蛋白则无此功能。结论:apo(a)kringleⅣ-10的Trp 72对其赖氨酸结合能力有关键作用。
Objective:To investigate the structural basis of lysine_binding heterogeneity of human Lipoprotein(a).Methods:Wild_type(wt)and mutant(mut)forms of apolipoprotein(a)[apo(a)]kringleⅣ-10 were expressed inE colistrain DH_5a as fusion GST_proteins by using recombinant plasmids pGEX_KG/Trp 72 and pGEX_KG/Arg 72,purified by glutathione_Agarose affinity chromatography.The lysine_binding capacities of the fusion proteins were examined by affinity chromatography on lysine_Sepharose.Results:The expressed GST_wt_kringleⅣ-10 fusion proteins bound avidly to lysine_Sepharose,but the expressed GST_mut_kringleⅣ-10 fusion proteins had no affinity for lysine_Sepharose.Conclusion:Apo(a)kringleⅣ-10 has lysine_binding capacities that requires the presence of Trp 72 and is independent of glycosylation.
作者
喻红
洪嘉玲
王韵
汪炳华
武军驻
Yu Hong;Hong Jialing;Wang Yun(Department of Biochemistry,BasicMedical College,HuLei Medicial University,Wuhan 430071,China)
基金
湖北省卫生厅资助课题No:97407
