期刊论文

中文

华中科技大学学报(医学版)

1672-0741

2004

33

5

526-530

10.3870/j.issn.1672-0741.2004.05.003

国家自然科学基金

本校为第一机构

目的研究VLDL-R中8个配体结合重复序列(ligand-binding repeats,LBR)在配体结合中的作用并探讨结合位点的结构.方法采用基因缺失诱变方法,构建不同LBR缺失的VLDL-R重组体.将其分别导入无LDL-R功能性表达的ldl-A7细胞.配体结合实验观察转染细胞与荧光标记的VLDL、β-VLDL的结合能力,并采用同源建模的方法预测受体N-端328个氨基酸的配体结合域的空间结构.结果配体结合实验显示LBR1和LBR2对受体结合VLDL、β-VLDL最为重要;LBR3对结合VLDL也有重要作用,但对结合β-VLDL无明显影响.模型显示受体N-端328个氨基酸的配体结合域呈现弧形口袋样结构,前3个LBR结构紧凑,呈棒状,负电荷相对集中,与功能特性吻合...

Objective To study the contribution of VLDL-R's 8 repeats to ligand-binding and to explore the structure of ligand-binding sites. Methods By using method of oligonucleotide-directed mutagenesis, VLDL-R recombinants lacking different repeats were constructed and transfected into ldl-A7 cells. The ligand binding assay was conducted with Dil labeled VLDL and (3-VLDL on the transfected cells. The method of protein structure prediction was adapted to analyze the 3D structure of the 328 amino acids sequence at the receptor's N-end. Results Ligand bin...