A site-directed mutagenesis study of drug-binding selectivity in genetic variants of human alpha(1)-acid glycoprotein.
A site-directed mutagenesis study of drug-binding selectivity in genetic variants of human alpha(1)-acid glycoprotein.
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DOI：
10.1002/jps.21697
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发表时间：
2009-11
影响因子：
3.8
通讯作者：
K. Nishi;M. Ueno;Y. Murakami;Naoko Fukunaga;T. Akuta;D. Kadowaki;Hiroshi Watanabe;A. Suenaga;T. Maruyama;M. Otagiri
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文献类型：
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作者：
K. Nishi;M. Ueno;Y. Murakami;Naoko Fukunaga;T. Akuta;D. Kadowaki;Hiroshi Watanabe;A. Suenaga;T. Maruyama;M. Otagiri
人α 1-酸性糖蛋白（AGP）是许多基础药物的主要载体，至少由两种遗传变异体组成，即A和F1*S变异体。有趣的是，AGP的变体具有不同的药物结合特性。本研究的目的是确定氨基酸残基，负责选择性的药物结合的遗传变异的AGP使用定点诱变。首先，我们筛选的氨基酸残基的位置100附近的区域，参与结合华法林和双嘧达莫，这是F1* S-特异性配体，普罗帕酮，这是一个A-特异性配体，使用超滤。在F1*S变体中，His 97、His 100和Trp 122参与华法林或双嘧达莫结合，而Glu 92、His 100和Trp 122参与A变体中普罗帕酮的结合。交换的残基之间的位置92 AGP变体逆转普罗帕酮结合的两个变体的相对强度，但有一个显着不同的效果结合华法林和潘生丁。这些发现表明，位置92处的氨基酸残基在AGP变体中的药物结合选择性中起重要作用，特别是对于优先结合A变体的药物。
Human alpha(1)-acid glycoprotein (AGP), a major carrier of many basic drugs in circulation, consists of at least two genetic variants, namely A and F1*S variant. Interestingly, the variants of AGP have different drug-binding properties. The purpose of this study was to identify the amino acid residues that are responsible for the selectivity of drug binding to genetic variants of AGP using site-directed mutagenesis. First, we screened amino acid residues in the region proximal to position 100 that are involved in binding of warfarin and dipyridamole, which are F1*S-specific ligands, and of propafenone, which is an A-specific ligand, using ultrafiltration. In the F1*S variant, His97, His100, and Trp122 were involved in either warfarin- or dipyridamole-binding, while Glu92, His100, and Trp122 participated in the binding of propafenone in the A variant. Exchange of the residue at position 92 between AGP variants reversed the relative strength of propafenone binding to the two variants, but had a markedly different effect on binding of warfarin and dipyridamole. These findings indicate that the amino acid residue at position 92 plays a significant role in drug-binding selectivity in AGP variants, especially for drugs that preferentially bind to the A variant.