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首页> 外文期刊>Biophysical Chemistry: An International Journal Devoted to the Physical Chemistry of Biological Phenomena >The location of the high- and low-affinity bilirubin-binding sites on serum albumin: Ligand-competition analysis investigated by circular dichroism
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The location of the high- and low-affinity bilirubin-binding sites on serum albumin: Ligand-competition analysis investigated by circular dichroism

机译：高和低亲和力胆红素结合位点在血清白蛋白上的位置：配体竞争分析的圆二色性

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摘要

The locations of three bilirubin (BR)-binding sites with different affinities were identified as subdomains IB, HA and IIIA for five mammalian serum albumins (SAs): human (HSA), bovine (BSA), rat, (RSA), rabbit (RbSA) and sheep (SSA). The stereoselectivity of a high-affinity BR-binding site was identified in the BR/ SA = 1/1 system by circular dichroism (CD) spectroscopy, the sites with low affinity to BR were analyzed using difference CD. Site-specific ligand-competition experiments with ibuprofen (marker for subdomain IIIA) and hemin (marker for subdomain IB) did not reveal any changes for the BR/SA = 1/1 system and showed a decrease of the bound BR at BR/SA = 3/1. Both sites were identified as sites with low affinity to BR. The correlation between stereoselectivity and the arrangement of Arg-Lys residues indicated similarity between the BR-binding sites in subdomain IIIA for all of the SAs studied. Subdomain IB in HSA, BSA, SSA and RbSA has P-stereoselectivity while in RSA it has M-selectivity toward BR. A ligand-competition experiment with gossypol shows a decrease of the CD signal of bound BR for the BR/SA =1/1 system as well as for BR/SA = 3/1. Subdomain IIA was assigned as a high-affinity BR-binding site.
机译：将三个具有不同亲和力的胆红素（BR）结合位点的位置确定为五个哺乳动物血清白蛋白（SA）的亚域IB，HA和IIIA：人（HSA），牛（BSA），大鼠（RSA），兔（ RbSA）和绵羊（SSA）。通过圆二色性（CD）光谱在BR / SA = 1/1系统中鉴定了高亲和力BR结合位点的立体选择性，使用差异CD分析了与BR的亲和力低的位点。用布洛芬（亚结构域IIIA的标记）和血红素（亚结构域IB的标记）进行的位点特异性配体竞争实验未揭示BR / SA = 1/1系统的任何变化，并且显示了BR / SA上结合的BR减少= 3/1。两个位点都被确定为与BR亲和力低的位点。立体选择性与Arg-Lys残基的排列之间的相关性表明，对于所有研究的SA，亚结构域IIIA中的BR结合位点之间都相似。 HSA，BSA，SSA和RbSA中的亚域IB具有P-立体选择性，而在RSA中，其对BR具有M-选择性。棉酚的配体竞争实验表明，对于BR / SA = 1/1系统以及BR / SA = 3/1，结合的BR的CD信号降低。将亚域IIA分配为高亲和力BR结合位点。

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