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从一种口腔密螺旋体中分离、鉴定其具有多边形阵列的外鞘并进行重组。

Isolation, properties, and reassembly of outer sheath carrying a polygonal array from an oral treponeme.

作者信息

Masuda K, Kawata T

出版信息

J Bacteriol. 1982 Jun;150(3):1405-13. doi: 10.1128/jb.150.3.1405-1413.1982.

DOI:10.1128/jb.150.3.1405-1413.1982
PMID:7076622
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC216367/
Abstract

The outer sheath carrying a polygonal array was isolated from an oral treponeme, Treponema sp. strain E-21, by disruption of cells by means of repeated freeze-thawing and by removal of flagella under acidic conditions followed by linear sucrose density gradient centrifugation. Electron microscopy revealed that the outer sheath was isolated as a triple-layered vesicle having a polygonal array, free of flagella and wall membrane complex. Using optical diffraction, negatively stained preparations of the outer sheath fragments showed that the polygonal array appeared to be composed of a hexagonal pattern with a predominant spacing of about 16.3 nm. The isolated outer sheath contained 49.7% protein, 30.8% total lipid, and 11.0% carbohydrate. Phospholipid comprised about 95% of the total lipid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the outer sheath was composed primarily of one major protein with an apparent molecular weight of about 62,000. The material from the isolated outer sheath solubilized with 1% sodium deoxycholate was reassembled into vesicles having a roughly polygonal array upon removal of the detergent by dialysis against 10 mM Tris-hydrochloride buffer with or without Mg2+.

摘要

通过反复冻融使细胞破裂，并在酸性条件下去除鞭毛，随后进行线性蔗糖密度梯度离心，从口腔密螺旋体（梅毒螺旋体属菌株E-21）中分离出带有多边形阵列的外鞘。电子显微镜显示，外鞘被分离为具有多边形阵列的三层囊泡，没有鞭毛和壁膜复合物。使用光学衍射，外鞘片段的负染制剂显示多边形阵列似乎由六边形图案组成，主要间距约为16.3nm。分离出的外鞘含有49.7%的蛋白质、30.8%的总脂质和11.0%的碳水化合物。磷脂约占总脂质的95%。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，外鞘主要由一种表观分子量约为62000的主要蛋白质组成。用1%脱氧胆酸钠溶解的分离外鞘材料，在通过与含或不含Mg2+的10mM Tris-盐酸缓冲液透析去除去污剂后，重新组装成具有大致多边形阵列的囊泡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/58098d046260/jbacter00259-0424-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/ea087cbccaf5/jbacter00259-0419-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/7ea7c4211a5f/jbacter00259-0420-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/4c9d621b911b/jbacter00259-0420-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/b39ea1e31154/jbacter00259-0422-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/f2904255d1ef/jbacter00259-0423-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/58098d046260/jbacter00259-0424-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/ea087cbccaf5/jbacter00259-0419-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/7ea7c4211a5f/jbacter00259-0420-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/4c9d621b911b/jbacter00259-0420-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/b39ea1e31154/jbacter00259-0422-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/f2904255d1ef/jbacter00259-0423-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d3/216367/58098d046260/jbacter00259-0424-a.jpg

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