New Structural Proteins of Halobacterium salinarum Gas Vesicle Revealed by Comparative Proteomics Analysis
New Structural Proteins of Halobacterium salinarum Gas Vesicle Revealed by Comparative Proteomics Analysis
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DOI：
10.1021/pr1009383
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发表时间：
2011-03-01
影响因子：
4.4
通讯作者：
Ng, Wailap Victor
中科院分区：
文献类型：
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作者：
Chu, Lichieh Julie;Chen, Mengchieh Claire;Ng, Wailap Victor
盐生盐杆菌气体囊泡（GV）是一种非常稳定的细胞内细胞器，空气被困在蛋白质膜内。本文报道了GV和GV耗竭裂解物（GVD）的比较蛋白质组学分析，以揭示膜结构蛋白。gvp-1编码的十种蛋白质（gvpMLKJIHGFED-1和gvpACNO-1）和由gvp-2编码的五种蛋白质通过LC-MS/MS鉴定了分别用于纺锤形和圆柱形GV的生物发生的（gvpMLKJIHGFED-2和gvpACNO-2）基因簇。GypA 1、I1、J1、A2和J2的肽仅在纯化的GV中鉴定，GypD 1、H1、L1和F2仅在GVD中鉴定，以及两个样品中的GypC 1、N1、O 1、F1、H2和O2。GypA 1、C1、F1、J1和A2在GV中的鉴定与它们先前已知的结构功能一致。此外，GypI 1、N1、O 1、H2、J2和O2在GV中的检测表明它们是新的结构蛋白。其中，GypI 1和N1在CV中的结构作用通过免疫检测纯化GV中蛋白A标记的GvpI 1和N1融合蛋白而进一步验证。因此，LC-MS/MS可以揭示至少六个气泡结构蛋白的主要纺锤形GV，可能有助于研究其生物成因。
The Halobacterium salinarum gas vesicle (GV) is an extremely stable intracellular organelle with air trapped inside a proteinaceous membrane. Reported here is a comparative proteomics analysis of GV and GV depleted lysate (GVD) to reveal the membrane structural proteins. Ten proteins encoded by gvp-1 (gvpMLKJIHGFED-1 and gvpACNO-1) and five proteins encoded by gvp-2 (gvpMLKJIHGFED-2 and gvpACNO-2) gene clusters for the biogenesis of spindle- and cylindrical-, respectively, shaped GV were identified by LC-MS/MS. The peptides of GypA1, I1, J1, A2, and J2 were exclusively identified in purified GV, GypD1, H1, L1, and F2 only in GVD, and GypC1, N1, O1, F1, H2, and O2 in both samples. The identification of GypA1, C1, F1, J1, and A2 in GV is in agreement with their previously known structural function. In addition, the detection of GypI1, N1, O1, H2, J2, and O2 in GV suggested they are new structural proteins. Among these, the structural role of GypI1 and N1 in CV was further validated by immuno-detection of protein A-tagged GvpI1 and N1 fusion proteins in purified GV. Thus, LC-MS/MS could reveal at least a half dozen gas vesicle structural proteins in the predominant spindle-shaped GV that may be helpful for studying its biogenesis.