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Structure and formation of PHB granule in bacteria

细菌中PHB颗粒的结构和形成

基本信息

批准号：
106658829
负责人：
Professor Dr. Dieter Jendrossek
金额：
--
依托单位：
Institut für Mikrobiologie (IMB)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2009
资助国家：
德国
起止时间：
2008-12-31 至 2014-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/106658829?language=en
关键词：
Structure formation PHB granule bacteria

项目摘要

Poly(3-hydroxybutyrate), PHB, is the most important storage compound for carbon and energy in prokaryotes. PHB granules at the early stages of formation are not randomly distributed in the cells but are – in contrast to literature – localized at the cell poles and/or at the inner side of the cytoplasmic membrane suggesting a specific interaction of PHB granule-associated proteins with unknown target proteins in the cell periphery. This assumption is supported by recent identification of cytoskeleton-like structures on the surface of isolated PHB granules by others. In this study subcellular localisation of active and inactive PHB synthase-Gfp fusion will be determined at different growth stages in Ralstonia eutropha (model organism for PHB accumulation, but small cell diameter) and Azotobacter vinelandii (PHB accumulation in cells with ø ≥2μm). A set of different fluorescent dyes with specificity for either cytoplasmic membrane, cytoplasm and PHB granules will be used to follow localization of PHB synthase-Gfp fusion and PHB granules formation in individual living cells. Proteins that specifically interact with PHB synthase and with phasin proteins (proteins of the PHB granule surface) will be identified by two hybrid analysis. Function of identified proteins will be investigated by disruption mutant analysis.

聚（3-羟基丁酸酯）是原核生物中最重要的碳和能量储存化合物。PHB颗粒在形成的早期阶段并不是随机分布在细胞中，而是（与文献相反）定位于细胞极点和/或细胞质膜的内侧，这表明PHB颗粒相关蛋白与细胞周围未知靶蛋白存在特异性相互作用。这一假设得到了最近其他人在分离的PHB颗粒表面发现的细胞骨架样结构的支持。在本研究中，将测定富营养菌Ralstonia europha （PHB积累的模式生物，但细胞直径较小）和vinelandii Azotobacter vinelandii （PHB在直径≥2μm的细胞中积累）在不同生长阶段活性和非活性PHB合成酶- gfp融合的亚细胞定位。一组对细胞质膜、细胞质和PHB颗粒具有特异性的不同荧光染料将用于追踪PHB合成酶- gfp融合和PHB颗粒在单个活细胞中的定位。与PHB合成酶和相蛋白（PHB颗粒表面的蛋白质）特异性相互作用的蛋白质将通过两次杂交分析确定。鉴定蛋白的功能将通过中断突变体分析进行研究。