Peptide binding domains determined through chemical modification of the side-chain functional groups.
Peptide binding domains determined through chemical modification of the side-chain functional groups.
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通过侧链官能团的化学修饰确定肽结合域。
DOI：
10.1016/s0006-3495(95)79934-9
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Houghten,RA
中科院分区：
文献类型：
--
作者：
Blondelle,SE;Perez-Paya,E;Allicotti,G;Forood,B;Houghten,RA
清楚地了解蛋白质和肽与脂质表面相互作用产生的特定二级结构和结合结构域将提供对生物活性分子的特定功能的洞察。我们已经表明，在早期的研究中，用于反相高效液相色谱的固定相代表一个模型人工脂质表面的诱导构象状态的肽对脂质相互作用的研究。我们现在已经使用反相高效液相色谱法，以确定结合域的肽，并通过扩展，蛋白质的脂质表面。该方法包括在肽与反相高效液相色谱C18基团相互作用后对特定氨基酸侧链官能团进行化学修饰。在相同的肽与脂质体结合后，还研究了对氧化的敏感性。选择通过两亲性α-螺旋18-mer肽的单个甲硫氨酸残基的氧化来说明这种方法。发现氧化程度明显取决于蛋氨酸残基对水移动的相的可及性。在其脂质诱导的构象状态的肽中发现的结合结构域是明确的两亲性α-螺旋的整个疏水面。
A clear understanding of the specific secondary structure and binding domain resulting from the interactions of proteins and peptides with lipid surfaces will provide insight into the specific functions of biologically active molecules. We have shown in earlier studies that the stationary phases used in reverse-phase high-performance liquid chromatography represent a model artificial lipid surface for the study of induced conformational states of peptides on lipid interaction. We have now used reverse-phase high-performance liquid chromatography to determine the binding domains of peptides and, by extension, of proteins to a lipid surface. This approach consists of performing chemical modifications of specific amino acid side-chain functionalities after the interaction of the peptides with the reverse-phase high-performance liquid chromatography C18 groups. The susceptibility to oxidation was also studied after binding of the same peptides to liposomes. Oxidation of a single methionine residue "walked" through an amphipathic alpha-helical 18-mer peptide was selected to illustrate this approach. The extent of oxidation was found to be clearly dictated by the accessibility of the methionine residue to the aqueous mobile phase. The binding domain found for the peptide in its lipid-induced conformational state was unequivocally the entire hydrophobic face of the amphipathic alpha-helix.