SOLUTION STRUCTURE AND FOLDING OF MYELIN BASIC PROTEIN

本研究将对牛的溶液结构和折叠进行研究 髓鞘碱性蛋白物种已被分离成 非磷化，一个磷酸盐残留物，两个磷酸盐残留物和三个 磷残留物种类。四种髓鞘碱性蛋白的每一种 电荷异构体将与交联剂发生反应 2-(对硝基苯基)烯丙基三甲基碘化铵和 2-(p-nitrophenyl)allyl-4-nitro-3-carboxyphenyl硫化物。这些试剂 通过Michael加成与赖氨酸残基反应，得到 热力学控制而不是其他方法产生的动力学控制 交联剂。分子内和分子间的交联蛋白 将被分离，所涉及的赖氨酸残留量将通过 胰酶多肽图谱和氨基酸分析。平行实验 乙二醇双(丁二酰亚胺)琥珀酸酯的合成 还将进行运动控制。关于交联剂的分析 结果应允许测定近端赖氨酸残留量。 折叠的蛋白质。磷酸盐残留物对环境质量的影响 将使用拉曼光谱研究蛋白质的二级结构和折叠。 傅立叶变换红外光谱。此外，还研究了pH值、离子强度和溶剂的影响 关于扩散系数和回转半径的确定 使用准弹性光散射。这项研究的结果将有助于 了解结构在该蛋白抗原性中的作用。在……里面 转变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。这些数据还将有助于 了解这种蛋白质在髓鞘膜中的作用。

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.