The effect of glycation on the structure, function and biological fate of human serum albumin as revealed by recombinant mutants
The effect of glycation on the structure, function and biological fate of human serum albumin as revealed by recombinant mutants
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DOI：
10.1016/j.bbagen.2003.08.001
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发表时间：
2003-10-13
影响因子：
3
通讯作者：
Otagiri, M
中科院分区：
文献类型：
--
作者：
Nakajou, K;Watanabe, H;Otagiri, M
利用酵母表达系统制备了重组野生型人血清白蛋白（rHSA）、单残基突变体K199A、K439A和K525A以及三残基突变体K199A/K439A/K525A。部分rHSA被糖化到不同程度（2.5-250 mM -葡萄糖）。通过远紫外和近紫外CD、固有色氨酸荧光和1,1'-双（4-苯胺）萘-5,5-二磺酸探针检测，单残基突变对白蛋白构象没有影响，而三残基突变和糖基化引起构象变化。三残基突变和糖基化对华法林高亲和力结合（位点I）的影响增加，但对丹氨酸肌氨酸高亲和力结合（位点II）和白蛋白酯酶样活性的影响降低。发现大鼠血浆半衰期与糖化rHSA （50 mM葡萄糖）有关。
Recombinant wild-type human serum albumin (rHSA), the single-residue mutants K199A, K439A and K525A and the triple-residue mutant K199A/K439A/K525A were produced using a yeast expression system. Portions of the rHSA were glycated to different degrees (2.5-250 mM -glucose). As detected by far-UV and near-UV CD, intrinsic tryptophan-fluorescence and probed by 1,1'-bis(4-anilino)naphthalene-5,5-disulfonic acid, the single-residue mutations had no effect on albumin conformation, whereas the triple-residue mutation and glycation caused conformational changes. The triple-residue mutation and glycation had comparable increased effects on high-affinity binding of warfarin (site I), but decreased effects on high-affinity binding of dansylsarcosine (site II) and the esterase-like activity of albumin. The relation between plasma half-lives in rats were found to be glycated rHSA (50 mM glucose)