Identification of cold-inducible inner membrane proteins of the psychrotrophic bacterium, Shewanella livingstonensis Ac10, by proteomic analysis
Identification of cold-inducible inner membrane proteins of the psychrotrophic bacterium, Shewanella livingstonensis Ac10, by proteomic analysis
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DOI：
10.1007/s00792-011-0422-z
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发表时间：
2012-01
期刊：
影响因子：
2.9
通讯作者：
Jungha Park;J. Kawamoto;N. Esaki;T. Kurihara
中科院分区：
文献类型：
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作者：
Jungha Park;J. Kawamoto;N. Esaki;T. Kurihara
希瓦氏菌（Shewanella livingstonensis Ac 10）是一种嗜冷的革兰氏阴性细菌，在接近0°C的温度下生长。该细菌的先前蛋白质组学研究鉴定了可能参与其冷适应的冷诱导可溶性蛋白和外膜蛋白（Kawamoto等人，Extremophiles 11：819-826，2007）。在这项研究中，我们建立了一种方法分离的内膜和外膜的蔗糖密度梯度超离心和内膜馏分进行蛋白质组学研究。将细胞在4 ° C和18°C的温度下生长，并获得富含磷脂的内膜级分。蛋白质的二维聚丙烯酰胺凝胶电泳和肽质量指纹分析鉴定了14种冷诱导蛋白（在4°C下增加超过2倍）。这些蛋白质中有六个被预测为内膜蛋白。两个预测的周质蛋白，5个预测的细胞质蛋白，和1个预测的外膜蛋白也被发现在内膜部分，这表明它们与内膜蛋白和/或脂质。这些冷诱导蛋白包括被认为参与趋化性（AtoS和PspA）、膜蛋白生物发生（DegP、SurA和FtsY）和形态发生（MreB）的蛋白。这些结果为进一步研究该菌的冷适应机制奠定了基础。
Shewanella livingstonensisAc10 is a psychrotrophic Gram-negative bacterium that grows at temperatures close to 0°C. Previous proteomic studies of this bacterium identified cold-inducible soluble proteins and outer membrane proteins that could possibly be involved in its cold adaptation (Kawamoto et al. in Extremophiles 11:819–826, 2007). In this study, we established a method for separating the inner and outer membranes by sucrose density gradient ultracentrifugation and performed proteomic studies of the inner membrane fraction. The cells were grown at temperatures of 4 and 18°C, and phospholipid-enriched inner membrane fractions were obtained. Two-dimensional polyacrylamide gel electrophoresis and peptide mass fingerprinting analysis of the proteins identified 14 cold-inducible proteins (more than a 2-fold increase at 4°C). Six of these proteins were predicted to be inner membrane proteins. Two predicted periplasmic proteins, 5 predicted cytoplasmic proteins, and 1 predicted outer membrane protein were also found in the inner membrane fraction, suggesting their association with the inner membrane proteins and/or lipids. These cold-inducible proteins included proteins that are presumed to be involved in chemotaxis (AtoS and PspA), membrane protein biogenesis (DegP, SurA, and FtsY), and morphogenesis (MreB). These findings provide a basis for further studies on the cold-adaptation mechanism of this bacterium.