SOLUTION STRUCTURE AND FOLDING OF MYELIN BASIC PROTEIN

本研究将研究牛的溶液结构和折叠 髓磷脂碱性蛋白种类已被分离为 非磷酸化，一个磷酸残基，两个磷酸残基和三个 磷酸盐残留物种类。 四种髓磷脂碱性蛋白中的每一种 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，得到产物 热力学控制而不是其他方法产生的动力学控制 交联剂。 分子内和分子间交联蛋白 将被分离，并且所涉及的赖氨酸残基将通过以下方式确定 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)得到产物 还将进行动力学控制。 交联分析 结果应允许确定近端赖氨酸残基 折叠的蛋白质。 磷酸盐残留量的影响 将使用拉曼研究蛋白质的二级结构和折叠 和 FT-IR 光谱。 此外，pH、离子强度和溶剂的影响 扩散系数和回转半径将被确定 使用准弹性光散射。 这项研究的结果将有助于 了解结构在该蛋白质抗原性中的作用。 在 反过来，这种理解将有助于帮助理解 多发性硬化症和 EAE 中的蛋白质。 该数据还将有助于 了解这种蛋白质在髓磷脂膜中的作用。

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.