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Mass spectrometry and NMR analysis of ligand binding by human liver fatty acid binding protein

机译:人肝脂肪酸结合蛋白结合配体的质谱和NMR分析

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摘要

Human liver fatty acid binding protein (hL-FABP) is the most abundant cytosolic protein in the liver. This protein plays important roles associated to partitioning of fatty acids (FAs) to specific metabolic pathways, nuclear signaling and protection against oxidative damage. The protein displays promiscuous binding properties and can bind two internal ligands, unlike FABPs from other tissues. Different topologies for the ligand located in the more accessible site have been reported, with either a 'head-in' or 'head-out' orientation of the carboxylate end. Electrospray-ionization mass spectrometry and nuclear magnetic resonance titrations are employed here in order to investigate in further detail the binding properties of this system, the equilibria established in solution and the pH dependence of the complexes. The results are consistent with two binding sites with different affinity and a unique head-out topology for the second molecule of either ligand. Competition experiments indicate a higher affinity for oleic acid relative to palmitic acid at each binding site.
机译:人肝脂肪酸结合蛋白(hL-FABP)是肝脏中最丰富的胞质蛋白。该蛋白起着重要的作用,与脂肪酸(FAs)分配到特定的代谢途径,核信号传导和防止氧化损伤有关。该蛋白表现出混杂的结合特性,可以结合两个内部配体,这与其他组织的FABP不同。已经报道了位于更易接近位点的配体的不同拓扑结构,其羧酸酯端具有“头朝内”或“头朝外”的取向。为了进一步详细研究该系统的结合特性,溶液中建立的平衡以及配合物的pH依赖性,此处采用电喷雾电离质谱法和核磁共振滴定法。结果与两个具有不同亲和力的结合位点和任一配体的第二个分子的独特抬头拓扑结构一致。竞争实验表明,在每个结合位点,相对于棕榈酸,对油酸的亲和力更高。

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