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SOLUTION STRUCTURE AND FOLDING OF MYELIN BASIC PROTEIN

髓磷脂碱性蛋白的溶液结构和折叠

基本信息

批准号：
3406995
负责人：
ROBERT ZAND
金额：
$ 7.01万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-04-01 至 1990-03-31
项目状态：
已结题

项目摘要

This study will investigate the solution structure and folding of bovine myelin basic protein species that have been separated into the nonphosphorylated, one phosphate residue, two phosphate residue and three phosphate residue species. Each one of the four myelin basic protein charge isomers will be reacted with the crosslinking agents 2-(p-nitrophenyl)allyltrimethylammonium iodide and 2-(p-nitrophenyl)allyl-4-nitro-3-carboxyphenyl sulfide. These reagents react with lysine residues via Michael addition to give products of thermodynamic control rather than kinetic control as produced by other crosslinking agents. The intra and inter molecularly crosslinked protein will be separated and the lysine residues involved will be determined via tryptic peptide mapping and amino acid analysis. Parallel experiments using ethylene glycol bis (succinimidyl succinate) to give the products of kinetic control will also be carried out. Analysis of the crosslinking results should permit a determination of the proximal lysine residues in the folded protein. The influence of the phosphate residues on the secondary structure and folding of the protein will be studied using Raman and FT-IR spectroscopy. Also, the effect of pH, ionic strength and solvent on the diffusion coefficient and radius of gyration will be determined using quasielastic light scattering. The results of this study will assist in understanding the role of structure in antigenicity of this protein. In turn that understanding will assist in helping to understand the role of the protein in Multiple Sclerosis and EAE. The data will also assist in understanding the role of this protein in the myelin membrane.

本研究将探讨牛血清白蛋白的溶液结构和折叠髓鞘碱性蛋白种类已被分离成非磷酸化，一个磷酸残基，两个磷酸残基和三个磷酸残基磷酸盐残留物。四种髓鞘碱性蛋白电荷异构体将与交联剂反应 2-（对硝基苯基）烯丙基三甲基碘化铵和 2-（对硝基苯基）烯丙基-4-硝基-3-羧基苯基硫醚。这些试剂通过迈克尔加成与赖氨酸残基反应，得到以下产物：热力学控制而不是动力学控制，交联剂分子内和分子间交联蛋白将被分离，所涉及的赖氨酸残基将通过胰蛋白酶肽图谱和氨基酸分析。平行实验使用乙二醇双（琥珀酰亚胺基琥珀酸酯）得到以下产物：还将进行动力学控制。交联分析结果应允许确定在细胞中的近端赖氨酸残基。折叠的蛋白质磷酸盐残留物对生物质降解的影响蛋白质的二级结构和折叠将使用拉曼进行研究和FT-IR光谱。此外，pH值，离子强度和溶剂的影响对扩散系数和回转半径的影响使用准弹性光散射。这项研究的结果将有助于在理解这种蛋白质的抗原性中结构的作用。在反过来，理解将有助于理解的作用，多发性硬化和EAE中的蛋白质。这些数据还将有助于了解这种蛋白质在髓鞘膜中的作用。