Binding of streptococcal lipoteichoic acid to the fatty acid binding sites on serum albumin.

Binding of streptococcal lipoteichoic acid to the fatty acid binding sites on serum albumin.
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链球菌脂磷壁酸与血清白蛋白上的脂肪酸结合位点的结合。

DOI:
10.1016/s0021-9258(18)43705-2
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发表时间:
1980
期刊:
The Journal of biological chemistry
影响因子:
--
通讯作者:
E. Beachey
E. Beachey
中科院分区:
--
文献类型:
--
作者:
W. A. Simpson;I. Ofek;E. Beachey

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研究了血清白蛋白的脂肪酸结合位点结合化脓链球菌脂磷壁酸的能力。初步研究表明,脂磷壁酸(而非其脱酰基剥夺剂)可以保护白蛋白免于因热(80℃,1 小时)而变性,并改变其在电场中的迁移率。与琼脂糖珠共价连接的白蛋白结合放射性标记的脂磷壁酸,并且结合的[3H]脂磷壁酸可以用未标记的脂磷壁酸或白蛋白特异性洗脱,但不能用其他测试的蛋白质洗脱。与白蛋白结合后,脂磷壁酸也可以用 50% 乙醇和各种去污剂定量洗脱,但不能用高达 1.0 M 的氯化钠。脂磷壁酸与白蛋白的结合遵循一级动力学,在 12 小时时达到饱和。通过斯卡查德图对结合数据的分析表明白蛋白分子上的结合位点的异质性类似于之前报道的脂肪酸。发现脂磷壁酸与白蛋白的结合亲和力分别介于先前报道的辛酸和棕榈酸之间。基于这些发现,我们制备了固定化白蛋白亲和柱,并能够从化脓性链球菌异质提取物中分离出具有生物活性的脂磷壁酸。
The ability of the fatty acid binding sites of serum albumin to bind lipoteichoic acid of Streptococcus pyogenes was investigated. Initial studies indicated that lipoteichoic acid, but not its deacylated deprivative, protected albumin from being denatured by heat (80C for 1 h) and changed its mobility in an electrical field. Albumin covalently linked to agarose beads bound radiolabeled lipoteichoic acid, and the bound [3H]lipoteichoic acid could be specifically eluted with unlabeled lipoteichoic acid or albumin but not with other proteins tested. After binding to albumins, the lipoteichoic acid also could be quantitatively eluted with 50% ethanol and various detergents but not with up to 1.0 M sodium chloride. Binding of lipoteichoic acid to albumin followed first order kinetics, reaching saturation at 12 h. Analysis of the binding data by a Scatchard plot indicated heterogeneity of the binding sites on the albumin molecule similar to that previously reported for fatty acids. The affinity of binding of lipoteichoic acid to albumin was found to be intermediate between that previously reported for octanoic and palmitic acids, respectively. Based on these findings, we prepared affinity columns of immobilized albumin and were able to separate biologically active lipoteichoic acid from heterogeneous extracts of S. pyogenes.