A lipopolysaccharide-binding cell-surface protein from Salmonella minnesota. Isolation, partial characterization and occurrence in different Enterobacteriaceae.
A lipopolysaccharide-binding cell-surface protein from Salmonella minnesota. Isolation, partial characterization and occurrence in different Enterobacteriaceae.
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来自明尼苏达沙门氏菌的脂多糖结合细胞表面蛋白。
DOI：
10.1111/j.1432-1033.1979.tb13156.x
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发表时间：
1979
期刊：
影响因子：
--
通讯作者：
J. Golecki
中科院分区：
文献类型：
--
作者：
R. Geyer;C. Galanos;O. Westphal;J. Golecki
1.通过用EDTA/NaCl溶液处理从沙门氏菌（Salmonella minillae Re）突变细胞获得的蛋白质提取物含有对细菌脂多糖表现出高亲和力的蛋白质。描述了这种脂多糖结合蛋白的分离和部分表征。2.从EDTA提取物中纯化的蛋白质由两个步骤的程序组成的离子交换色谱CM-Sephadex和制备型聚丙烯酰胺凝胶电泳在pH 9.5。总纯化步骤的产率约为16%。3.所得到的蛋白制备物是均匀的圆盘凝胶电泳，十二烷基硫酸盐凝胶电泳，聚丙烯酰胺凝胶等电聚焦和免疫电泳的基础上。4.发现该蛋白质的等电点在4 ℃时为10.3。十二烷基硫酸盐凝胶电泳测定其分子量为15000。其氨基酸组成的特征是不含组氨酸和脯氨酸，酪氨酸含量低，丙氨酸、赖氨酸、天冬氨酸和谷氨酸残基或其各自的酰胺含量高。5.脂多糖-蛋白质协会被证明主要是由于离子相互作用的基本蛋白质与带负电荷的基团（可能是磷酸盐和焦磷酸盐基团）的脂质A部分。6.纯化的脂多糖结合蛋白在兔中具有免疫原性，从而能够制备特异性抗血清。7.该蛋白质位于沙门氏菌的mineriform Re突变细胞的表面，如通过与总细菌的抗血清吸收所揭示的。铁蛋白标记研究进一步证明，它均匀地分布在整个细胞表面。8.使用明尼苏达沙门氏菌、鼠伤寒沙门氏菌、大肠杆菌、克雷伯氏菌和志贺氏菌的光滑和粗糙菌株进行的比较抗血清吸收研究表明，大多数测试菌株中都存在脂多糖结合蛋白（或血清学交叉反应抗原）。从这些结果可以认为该蛋白是肠杆菌科的共同抗原。
1. Protein extracts obtained from Salmonella minnesota Re mutant cells by treatment with EDTA/NaC1 solution contain a protein which exhibits high affinity to bacterial lipopolysaccharides. The isolation and partial characterization of this lipopolysaccharide-binding protein is described. 2. The protein was purified from EDTA extracts by a two-step procedure consisting of ion-exchange chromatography on CM-Sephadex and preparative polyacrylamide gel electrophoresis at pH 9.5. The yield of the total purification procedure was around 16%. 3. The resulting protein preparation was homogeneous on the basis of disc gel electrophoresis, dodecylsulfate gel electrophoresis, isoelectric focusing in polyacrylamide gel and immunoelectrophoresis. 4. The isoelectric point of the protein was found to be 10.3 at 4 degrees C. Its molecular weight determined by dodecylsulfate gel electrophoresis is 15000. Its amino acid composition is characterized by the absence of histidine and proline, a low content in tyrosine and high amounts of alanine, lysine, aspartic and glutamic acid residues, or their respective amides. 5. The lipopolysaccharide-protein association was shown to be mainly due to ionic interactions of the basic protein with negatively charged groups (probably phosphate and pyrophosphate groups) of the lipid A moiety. 6. Purified lipopolysaccharide-binding protein is immunogenic in rabbits, thus enabling the preparation of specific antiserum. 7. The protein is located at the surface of Salmonella minnesota Re mutant cells as revealed by antiserum absorption with total bacteria. Ferritin-labelling studies further demonstrated that it is evenly spread over the entire cell surface. 8. Comparative antiserum absorption studies using smooth and rough strains of Salmonella minnesota, Salmonella typhimurium, Escherichia coli, Klebsiella and Shigella revealed the presence of lipopolysaccharide-binding protein (or a serologically cross-reacting antigen) in most of the strains tested. From these results the protein can be considered as a common antigen of Enterobacteriaceae.