OUTER MEMBRANES OF GRAM-NEGATIVE BACTERIA .19. ISOLATION FROM PSEUDOMONAS-AERUGINOSA PAO1 AND USE IN RECONSTITUTION AND DEFINITION OF PERMEABILITY BARRIER
OUTER MEMBRANES OF GRAM-NEGATIVE BACTERIA .19. ISOLATION FROM PSEUDOMONAS-AERUGINOSA PAO1 AND USE IN RECONSTITUTION AND DEFINITION OF PERMEABILITY BARRIER
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DOI：
10.1128/jb.136.1.381-390.1978
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发表时间：
1978-01-01
影响因子：
3.2
通讯作者：
NIKAIDO, H
中科院分区：
文献类型：
--
作者：
HANCOCK, REW;NIKAIDO, H
设计了一种在不添加EDTA的情况下分离铜绿假单胞菌PAO1外膜和内膜的方法。该方法得到2个外膜组分，在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图上显示相同的蛋白质图谱，但在磷脂的相对含量上有很大差异。根据典型的内膜和外膜标志物的含量判断，其中一个外膜组分和内膜组分的交叉污染程度不到4%。外膜有4条主要蛋白带，表观分子量分别为37,000，35,000，21,000和17,000。在囊泡形成过程中，由脂多糖和磷脂组成的囊泡对囊泡中的所有糖类都是不渗透的。当囊泡含有外膜蛋白时，它们只完全保留那些大于9000 mW的糖，这表明铜绿假单胞菌外膜对糖的排除限度大大大于某些肠道细菌的数字(500-600道尔顿)。讨论了对亲水性物质具有较高排除极限的外膜的优点和潜在的缺点。
A method for separating the outer and inner membranes of P. aeruginosa PAO1 in the absence of added EDTA was devised. The method yields 2 outer membrane fractions which show the same protein pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but differ substantially in their relative contents of phospholipids. One of these outer membrane fractions and the inner membrane fraction are less than 4% cross-contaminated, as judged by the content of typical inner and outer membrane markers. The outer membrane contains 4 major protein bands with apparent MW of 37,000, 35,000, 21,000 and 17,000. Vesicles reconstituted from lipopolysaccharide and phospholipids were impermeable to all saccharides included in the vesicles during vesicle formation. When the vesicles contained outer membrane proteins, they fully retained only those saccharides of greater than 9000 MW, suggesting that the exclusion limit of the outer membrane of P. aeruginosa for saccharides is substantially larger than the figure (500-600 daltons) obtained for certain enteric bacteria. The advantages and potential disadvantages of having an outer membrane with a higher exclusion limit for hydrophilic substances are discussed.