脂肪酸结合蛋白的研究。大鼠肝脏脂肪酸结合蛋白的纯化及半胱氨酸-69在脂肪酸结合中的作用。
Studies on fatty-acid-binding proteins. The purification of rat liver fatty-acid-binding protein and the role of cysteine-69 in fatty acid binding.
作者信息
Wilton D C
机构信息
Department of Biochemistry, School of Biochemical and Physiological Sciences, University of Southampton, U.K.
出版信息
Biochem J. 1989 Jul 1;261(1):273-6. doi: 10.1042/bj2610273.
Abstract
- A new, simple and high-yield procedure is described for the purification of hepatic fatty-acid-binding protein from rat liver using naphthylaminodecyl-agarose as an affinity column. 2. Cysteine-69 is shown to react slowly, but quantitatively, with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), indicating that the thiol group is free, but may be buried within the protein. 3. Fatty acids do not affect the DTNB reactivity of this cysteine residue; however, cysteine reactivity is enhanced in the presence of haem and oleoyl-CoA. 4. Fatty-acid-binding protein that has been modified with DTNB is still able to bind the fluorescent fatty acid 11-(dansylamino)undecanoic acid, indicating that cysteine-69 may be remote from the fatty-acid-binding site.
摘要
- 描述了一种新的、简单且高产率的方法，该方法使用萘胺癸基琼脂糖作为亲和柱从大鼠肝脏中纯化肝脂肪酸结合蛋白。2. 结果表明，半胱氨酸-69与5,5'-二硫代双-(2-硝基苯甲酸)(DTNB)反应缓慢，但定量反应，这表明硫醇基团是游离的，但可能埋藏在蛋白质内部。3. 脂肪酸不影响该半胱氨酸残基的DTNB反应性；然而，在血红素和油酰辅酶A存在的情况下，半胱氨酸反应性增强。4. 用DTNB修饰的脂肪酸结合蛋白仍能结合荧光脂肪酸11-(丹磺酰氨基)十一烷酸，这表明半胱氨酸-69可能远离脂肪酸结合位点。
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