论文部分内容阅读
通过酵母重组P.rβ_2-GPI-DV(β2糖蛋白I第5结构域)蛋白的荧光、圆二色谱及分子对接模拟,分析血浆糖蛋白β2-GPI-DV分子结构域与ox-LDL的结合机制。SDS-PAGE、Western blotting验证重组蛋白P.rβ_2-GPI-DV表达正确性及纯度;荧光、圆二色光谱、分子对接模拟解析重组蛋白与oxLDL、oxLDL配体oxLig-1可能存在的结合机制和位点。SDS-PAGE、Western blotting显示,在12 kDa大小处有目的蛋白存在且与特异性抗体结合;荧光、圆二色谱的发色基团、二级结构的变化趋势及分子对接模拟结果揭示,P.rβ_2-GPI-DV的Cys281-Lys-Asn-Lys-Glu-Lys-Lys287和Leu313-Ala-Phe-Trp316区域组成的活性口袋与oxLig-1的ω-COOH羧基是实现二者结合的关键。以上结果表明,β2-GPI通过其第5结构域(DV)的赖氨酸阳性区域与ox-LDL的ω-COOH基团识别结合,为血清中β2-GPI特异性结合ox-LDL的机制研究提供理论依据。
The binding of plasma glycoprotein β2-GPI-DV molecular domain to ox-LDL was analyzed by fluorescence, circular dichroism and molecular docking simulation of the yeast recombinant P.rβ_2-GPI-DV (β2 glycoprotein I domain 5) protein mechanism. SDS-PAGE and Western blotting were used to verify the correctness and purity of recombinant protein P.rβ_2-GPI-DV. Fluorescence and circular dichroism spectroscopy and molecular docking were used to analyze the possible binding mechanism between recombinant protein and oxLDL and oxLDL-1 Site. SDS-PAGE and Western blotting showed that the target protein was present at 12 kDa and bound to the specific antibody. Fluorescence and circular dichroism of the chromogenic groups, the secondary structure and the molecular docking simulation revealed that, The active pocket composed of the Cys281-Lys-Asn-Lys-Glu-Lys-Lys287 and Leu313-Ala-Phe-Trp316 regions of rβ_2-GPI-DV and the ω-COOH carboxyl group of oxLig-1 are the key to the combination of the two. The above results indicate that β2-GPI can specifically bind ox-LDL to β2-GPI in serum through the recognition of the lysine-positive region of the fifth domain (DV) and the ω-COOH group of ox-LDL Provide a theoretical basis.