Localization of proteins in the inner and outer membranes of Caulobacter crescentus.
Localization of proteins in the inner and outer membranes of Caulobacter crescentus.
复制标题
新月柄杆菌内膜和外膜中蛋白质的定位。
DOI：
10.1016/0005-2736(82)90108-0
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Newton,A
中科院分区：
文献类型：
--
作者：
Clancy,MJ;Newton,A
通过等密度蔗糖梯度离心将新月柄杆菌的细胞质和外膜分离成两个峰，浮力密度分别为1.22和1.14 g/cm3。通过较低密度峰中苹果酸脱氢酶和 NADH 氧化酶的富集以及较重峰中鞭毛蛋白（一种细胞表面蛋白）的存在，将这些峰鉴定为外膜和细胞质膜。通过用重氮化[35S]磺胺酸（一种不能穿透完整细胞的试剂）标记细胞来确认较重的峰为外膜。在这些条件下，只有外膜蛋白被磺胺酸取代。通过SDS-聚丙烯酰胺和二维凝胶电泳分析[35S]甲硫氨酸标记的膜来检查细胞质和外膜之间的蛋白质分布。这些结果表明，内膜和外膜含有大致相等数量的蛋白质，并且这些蛋白质在两层之间的分布是高度不对称的。尽管许多蛋白质可以分配到一种或另一种膜组分，但许多分子量范围为 32 000-100 000 的外膜蛋白质经常污染内膜组分。这些结果对 C 中膜分离和分离的影响。讨论了新月形。
Cytoplasmic and outer membranes ofCaulobacter crescentuswere separated by isopycnic sucrose gradient centrifugation into two peaks with buoyant densities 1.22 and 1.14 g/cm3. These peaks were identified as outer and cytoplasmic membranes by the enrichment of malate dehydrogenase and NADH oxidase in the lower density peak and the presence of flagellin, a cell surface protein, in the heavier peak. The identity of the heavier peak as outer membrane was confirmed by labeling of cells with diazotized [35S]sulfanilic acid, a reagent that does not penetrate intact cells. Under these conditions only outer membrane proteins were substituted by the sulfanilic acid. The distribution of proteins between the cytoplasmic and outer membranes were examined by the analysis of [35S]methionine-labeled membranes by SDS-polyacrylamide and two-dimensional gel electrophoresis. These results showed that the inner and outer membranes contain approximately equal numbers of proteins, and that the distribution of these proteins between the two layers is highly asymmetric. Although many of the proteins could be assigned to one or the other membrane fraction, a number of the outer membrane proteins in the 32 000–100 000 molecular weight range frequently contaminate the inner membrane fractions. The implications of these results for membrane isolation and separation inC. crescentusare discussed.
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影响因子：
3.2
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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3.2
作者：
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新月柄杆菌发育过程中蛋白质合成的模式。
影响因子：
2.7
作者：
K. Cheung;A. Newton
通讯作者：
A. Newton
普瓦泽基立克次体内膜和外膜的分离及其多肽组成的表征
影响因子：
3.2
作者：
D. Smith;H. Winkler
通讯作者：
H. Winkler