载脂蛋白A-I(apolipoprotein A-I,ApoA-Ⅰ)是高密度脂蛋白(high density lipoprotein,HDL)的主要蛋白成分,在胆固醇逆转运(reverse cholesterol transport,RCT)中发挥关键作用,是抗动脉粥样硬化(atherosclerosis,As)药物研发的重要靶标...载脂蛋白A-I(apolipoprotein A-I,ApoA-Ⅰ)是高密度脂蛋白(high density lipoprotein,HDL)的主要蛋白成分,在胆固醇逆转运(reverse cholesterol transport,RCT)中发挥关键作用,是抗动脉粥样硬化(atherosclerosis,As)药物研发的重要靶标。靶向ApoA-Ⅰα螺旋研发的模拟肽和基于重组ApoA-Ⅰ研发的HDL模拟肽已进入临床试验,展示了良好的促RCT与抗As活性。本文综述了ApoA-Ⅰ的结构和功能,ApoA-Ⅰ与ABCA1(ATP-binding cassette transporter A1)、LCAT(lecithin cholesterol acyl transferase)、SR-B1(scavenger receptor class B type 1)的分子互作机制,总结了靶向ApoA-Ⅰ模拟肽的研究进展,以期为模拟肽类抗As新药研发提供参考。展开更多
To identify, clone ,sequence and highly express the mature peptide gene of ApoA Ⅰ, total RNA was prepared from human fetal liver tissue. cDNA fragment encoding human ApoA Ⅰ was amplified by RT-PCR using specific pri...To identify, clone ,sequence and highly express the mature peptide gene of ApoA Ⅰ, total RNA was prepared from human fetal liver tissue. cDNA fragment encoding human ApoA Ⅰ was amplified by RT-PCR using specific primers, and then was inserted in pGEM-T vector. DNA sequencing indicates that the fragment is 729 base pairs in length and has 100% nucleotide homology with that of reported ApoA Ⅰ cDNA gene previously. The ApoA Ⅰ gene was cloned into pGEX 5X-1.The recombinant protein was expressed in E.coli DH5α, purified by glutathione-Sepharose 4B affinity chromatography and confirmed by SDS-PAGE. It was shown that the recombinant ApoA Ⅰ was expressed in E.coli, and the target protein amounted to 36% of total bacteria proteins. Cholesteryl ester transfer experiment showed that the recombinant ApoA Ⅰ was capable of promoting transfer of CE from HDL to LDL. Western blotting showed that the protein could react specifically with anti-ApoA Ⅰ antibodies.展开更多
文摘载脂蛋白A-I(apolipoprotein A-I,ApoA-Ⅰ)是高密度脂蛋白(high density lipoprotein,HDL)的主要蛋白成分,在胆固醇逆转运(reverse cholesterol transport,RCT)中发挥关键作用,是抗动脉粥样硬化(atherosclerosis,As)药物研发的重要靶标。靶向ApoA-Ⅰα螺旋研发的模拟肽和基于重组ApoA-Ⅰ研发的HDL模拟肽已进入临床试验,展示了良好的促RCT与抗As活性。本文综述了ApoA-Ⅰ的结构和功能,ApoA-Ⅰ与ABCA1(ATP-binding cassette transporter A1)、LCAT(lecithin cholesterol acyl transferase)、SR-B1(scavenger receptor class B type 1)的分子互作机制,总结了靶向ApoA-Ⅰ模拟肽的研究进展,以期为模拟肽类抗As新药研发提供参考。
文摘To identify, clone ,sequence and highly express the mature peptide gene of ApoA Ⅰ, total RNA was prepared from human fetal liver tissue. cDNA fragment encoding human ApoA Ⅰ was amplified by RT-PCR using specific primers, and then was inserted in pGEM-T vector. DNA sequencing indicates that the fragment is 729 base pairs in length and has 100% nucleotide homology with that of reported ApoA Ⅰ cDNA gene previously. The ApoA Ⅰ gene was cloned into pGEX 5X-1.The recombinant protein was expressed in E.coli DH5α, purified by glutathione-Sepharose 4B affinity chromatography and confirmed by SDS-PAGE. It was shown that the recombinant ApoA Ⅰ was expressed in E.coli, and the target protein amounted to 36% of total bacteria proteins. Cholesteryl ester transfer experiment showed that the recombinant ApoA Ⅰ was capable of promoting transfer of CE from HDL to LDL. Western blotting showed that the protein could react specifically with anti-ApoA Ⅰ antibodies.
