Caamaño C A, Zand R
Biophysics Research Division, University of Michigan, Ann Arbor 48109-2099.
Biochem Int. 1989 Jun;18(6):1245-51.
The use of chemical crosslinking methodologies for the study of the solution structure and folding of the myelin basic protein required the development of a specific protocol for separating the various reaction products. Myelin basic protein treated with the crosslinking reagent dithiobis(succinimidylpropionate) was subjected to analysis by urea-SDS polyacrylamide gel electrophoresis. This permitted the identification of dimer and higher oligomeric crosslinked products. The dissociating conditions of this method precluded the dimerization of the basic protein observed in systems with SDS and without urea. Similar samples analyzed by gel filtration-fast protein liquid chromatography exhibited a complex elution pattern in contrast to the protein not reacted with the crosslinker. The electrophoretic analysis of the different eluted fractions revealed that at least three monomeric forms of modified myelin basic protein had been separated by gel filtration.
使用化学交联方法研究髓鞘碱性蛋白的溶液结构和折叠需要开发一种用于分离各种反应产物的特定方案。用交联剂二硫代双（琥珀酰亚胺丙酸酯）处理的髓鞘碱性蛋白通过尿素-SDS聚丙烯酰胺凝胶电泳进行分析。这使得能够鉴定二聚体和更高寡聚体的交联产物。该方法的解离条件排除了在含有SDS但不含尿素的系统中观察到的碱性蛋白的二聚化。与未与交联剂反应的蛋白质相比，通过凝胶过滤-快速蛋白质液相色谱分析的类似样品表现出复杂的洗脱模式。对不同洗脱级分的电泳分析表明，通过凝胶过滤至少分离出了三种修饰髓鞘碱性蛋白的单体形式。
