用正辛基-β-D-吡喃葡萄糖苷溶解晶状体纤维质膜的主要多肽（MP26）的高效液相色谱分析

High-performance liquid chromatography of the main polypeptide (MP26) of lens fiber plasma membranes solubilized with n-octyl beta-D-glucopyranoside.

作者信息

Manenti S, Dunia I, le Maire M, Benedetti E L

机构信息

Institut Jacques Monod, Centre National de la Recherche Scientifique (CNRS), Université Paris VII, France.

出版信息

FEBS Lett. 1988 Jun 6;233(1):148-52. doi: 10.1016/0014-5793(88)81373-5.

DOI:10.1016/0014-5793(88)81373-5

PMID:3164277

Abstract

The main polypeptide isolated from lens fiber membrane has been solubilized in octyl glucoside and studied by gel filtration in high-performance liquid chromatography (HPLC). The combination of S20,w value obtained from analytical ultracentrifugation and Stokes radius determined by HPLC of the soluble fraction indicates that more than 90% of the protein is monomeric. The solubilization of the protein seems to be dependent upon the presence of the NH2 and COOH terminal sequences, since proteolytic degradation of MP26 which removes these terminal sequences is less soluble than the uncleaved polypeptide. Moreover, there is a higher amount of oligomer after proteolysis. Fatty acid analysis by gas chromatography shows that the insoluble membrane fraction from both cortical and nuclear fibers comprises a special class of long (C22) saturated fatty acids (behenic acid).

摘要

从晶状体纤维膜中分离出的主要多肽已溶解在辛基葡糖苷中，并通过高效液相色谱（HPLC）中的凝胶过滤进行研究。从分析超速离心获得的S20,w值与通过HPLC测定的可溶性部分的斯托克斯半径相结合表明，超过90%的蛋白质是单体。蛋白质的溶解似乎取决于NH2和COOH末端序列的存在，因为去除这些末端序列的MP26的蛋白水解降解产物比未切割的多肽更难溶解。此外，蛋白水解后寡聚物的含量更高。通过气相色谱进行的脂肪酸分析表明，皮质和核纤维的不溶性膜部分都包含一类特殊的长链（C22）饱和脂肪酸（山嵛酸）。