Purification and characteristics of the streptococcal chemotactic factor inactivator.
Purification and characteristics of the streptococcal chemotactic factor inactivator.
复制标题
链球菌趋化因子灭活剂的纯化及特性。
DOI：
10.1128/iai.50.3.757-764.1985
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发表时间：
1985
影响因子：
3.1
通讯作者：
Cleary,PP
中科院分区：
文献类型：
--
作者：
Wexler,DE;Cleary,PP
研究了A群链球菌趋化因子灭活剂（SCFI）的生化和免疫学特性。SCFI通过有限胰蛋白酶消化从完整的M+细菌中提取，并通过硫酸铵分离、疏水相互作用层析和阴离子交换层析进行顺序纯化。凝胶渗透色谱和DEAE高压液相色谱分析表明，SCFI活性与多种物种有关。柱纯化蛋白的聚丙烯酰胺凝胶电泳也显示出高度的分子异质性，大部分材料在103,000至114,000 Mr的簇中。SCFI活性对多种蛋白水解酶的破坏很敏感，SCFI多克隆抗血清能够中和其抗趋化活性。SCFI的二维免疫电泳表明，各种共纯化物种具有免疫交叉反应性，并且在制备中具有高度的抗原同质性。对M+细菌的粗洗涤液提取物进行Western blot分析，发现一个主要的抗原种类对应于135,000 Mr和一个较少丰富的137,000 Mr. SCFI抗血清与M蛋白没有反应，因此SCFI似乎是一个独特的分子，尽管它与M+表型密切相关。
The biochemical and immunological characteristics of the chemotactic factor inactivator of group A streptococci (SCFI) were examined. SCFI was extracted from intact M+ bacteria by limited trypsin digestion and purified sequentially by ammonium sulfate fractionation, hydrophobic interaction chromatography, and anion-exchange chromatography. SCFI activity was associated with multiple species as indicated by gel permeation chromatography and DEAE high-pressure liquid chromatography analyses. Polyacrylamide gel electrophoresis of the column-purified protein also demonstrated a high degree of molecular heterogeneity, with most of the material in a 103,000 to 114,000 Mr cluster. SCFI activity was sensitive to destruction by several proteolytic enzymes, and polyclonal antiserum to SCFI was able to neutralize its antichemotactic activity. Two-dimensional immunoelectrophoresis of SCFI indicated that the various copurified species were immunologically cross-reactive and indicated a high degree of antigenic homogeneity within the preparation. Western blot analysis of crude detergent extracts of M+ bacteria identified a major antigenic species corresponding to 135,000 Mr and a less abundant species of 137,000 Mr. SCFI antiserum was not reactive with M protein, and therefore SCFI appeared to be a distinct molecule, despite its close association with the M+ phenotype.