Isolation, Composition, and Structure of Membrane of Listeria monocytogenes
Isolation, Composition, and Structure of Membrane of Listeria monocytogenes
复制标题
单核细胞增生李斯特氏菌的分离、组成和膜结构
DOI：
10.1128/jb.95.2.688-699.1968
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发表时间：
1968
影响因子：
3.2
通讯作者：
K. Carroll
中科院分区：
文献类型：
--
作者：
B. K. Ghosh;K. Carroll
单核细胞增生李斯特氏菌菌株 42 的质膜是通过用含有 MgCl2 和葡萄糖的三（羟甲基）氨基甲烷 (Tris) 缓冲液（pH 8.2）渗透裂解原生质体，然后用 Tris 缓冲液中的 NaCl 和 MgCl2 洗涤来制备的。电子显微镜显示该制剂没有被细胞质物质污染。膜制剂由55%至60%蛋白质、1.5%核糖核酸、0.1%脱氧核糖核酸、1.3%至2.3%碳水化合物、0.17%至0.38%氨基糖、0.2%至0.4%鼠李糖、3.5%至4.0%磷、10.5%至12.0%氮和30% 35%脂质。洗过的膜的氨基酸组成与整个细胞的氨基酸组成有一些差异。膜水解产物中不存在含硫氨基酸。膜碳水化合物含有葡萄糖、半乳糖、核糖和阿拉伯糖。膜脂为80～85％磷脂和15～20％中性脂质。该脂质含有2.3%至3.0%的磷、2.5%至3.0%的碳水化合物和极少量的氮（0.2%至0.3%）。磷脂是磷脂酰甘油类型。洗过的膜的电子显微照片显示出三层。外层和内层的厚度从25到37埃不等，中层从20到25埃不等。总厚度在85到100埃之间不等。这些制剂含有许多被柠檬酸铅严重染色的囊泡。一些囊泡还以挤压或侵入或两者的形式附着在原生质体上。在不存在 MgCl2 的情况下通过裂解原生质体获得的膜制剂比用 MgCl2 制备的制剂破碎，并且含有更少的脂质（20 至 22%）和核糖核酸（0.3 至 0.5%）。
The plasma membrane of Listeria monocytogenes strain 42 was prepared by osmotic lysis of protoplasts with tris(hydroxymethyl)aminomethane (Tris) buffer, pH 8.2, containing MgCl2 and glucose, followed by washing with NaCl and MgCl2 in Tris buffer. Electron microscopy showed that the preparation was not contaminated with cytoplasmic material. The membrane preparation was composed of 55 to 60% protein, 1.5% ribonucleic acid, 0.1% deoxyribonucleic acid, 1.3 to 2.3% carbohydrate, 0.17 to 0.38% amino sugar, 0.2 to 0.4% rhamnose, 3.5 to 4.0% phosphorus, 10.5 to 12.0% nitrogen, and 30 to 35% lipid. Amino acid composition of the washed membrane showed some variation from that of the whole cells. Sulfur-containing amino acids were not present in the membrane hydrolysate. The membrane carbohydrate contained glucose, galactose, ribose, and arabinose. The membrane lipid was 80 to 85% phospholipid and 15 to 20% neutral lipid. The lipid contained 2.3 to 3.0% phosphorus, 2.5 to 3.0% carbohydrate, and a very small amount of nitrogen (0.2 to 0.3%). The phospholipid was of the phosphatidyl glycerol type. Electron micrographs of the washed membrane showed three layers. The outer and inner layers varied in thickness from 25 to 37 A and the middle layer from 20 to 25 A. The total thickness varied between 85 and 100 A. These preparations contained many vesicles which stained heavily with lead citrate. Some vesicles were also attached to the protoplast ghosts in the form of extrusions or intrusions, or both. Membrane preparations obtained by lysis of protoplasts in the absence of MgCl2 were fragmented and contained less lipid (20 to 22%) and ribonucleic acid (0.3 to 0.5%) than preparations prepared with MgCl2.