Lai Erh-Min, Phadke Nikhil D, Kachman Maureen T, Giorno Rebecca, Vazquez Santiago, Vazquez Jenny A, Maddock Janine R, Driks Adam
Department of Molecular, Cellular and Developmental Biology, University of Michigan. Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
J Bacteriol. 2003 Feb;185(4):1443-54. doi: 10.1128/JB.185.4.1443-1454.2003.
The outermost proteinaceous layer of bacterial spores, called the coat, is critical for spore survival, germination, and, for pathogenic spores, disease. To identify novel spore coat proteins, we have carried out a preliminary proteomic analysis of Bacillus subtilis and Bacillus anthracis spores, using a combination of standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation and improved two-dimensional electrophoretic separations, followed by matrix-assisted laser desorption ionization-time of flight and/or dual mass spectrometry. We identified 38 B. subtilis spore proteins, 12 of which are known coat proteins. We propose that, of the novel proteins, YtaA, YvdP, and YnzH are bona fide coat proteins, and we have renamed them CotI, CotQ, and CotU, respectively. In addition, we initiated a study of coat proteins in B. anthracis and identified 11 spore proteins, 6 of which are candidate coat or exosporium proteins. We also queried the unfinished B. anthracis genome for potential coat proteins. Our analysis suggests that the B. subtilis and B. anthracis coats have roughly similar numbers of proteins and that a core group of coat protein species is shared between these organisms, including the major morphogenetic proteins. Nonetheless, a significant number of coat proteins are probably unique to each species. These results should accelerate efforts to develop B. anthracis detection methods and understand the ecological role of the coat.
细菌芽孢最外层的蛋白质层称为芽孢衣，对芽孢的存活、萌发以及对于致病性芽孢而言，对引发疾病都至关重要。为了鉴定新型芽孢衣蛋白，我们结合标准的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分离和改进的二维电泳分离方法，随后进行基质辅助激光解吸电离飞行时间质谱和/或串联质谱，对枯草芽孢杆菌和炭疽芽孢杆菌的芽孢进行了初步的蛋白质组学分析。我们鉴定出38种枯草芽孢杆菌芽孢蛋白，其中12种是已知的芽孢衣蛋白。我们提出，在这些新蛋白中，YtaA、YvdP和YnzH是真正的芽孢衣蛋白，我们分别将它们重新命名为CotI、CotQ和CotU。此外，我们启动了对炭疽芽孢杆菌芽孢衣蛋白的研究，鉴定出11种芽孢蛋白，其中6种是候选芽孢衣或芽孢外壁蛋白。我们还在未完成的炭疽芽孢杆菌基因组中查询潜在的芽孢衣蛋白。我们的分析表明，枯草芽孢杆菌和炭疽芽孢杆菌的芽孢衣蛋白数量大致相似，并且这些生物体之间共享一组核心的芽孢衣蛋白种类，包括主要的形态发生蛋白。尽管如此，每个物种可能都有大量独特的芽孢衣蛋白。这些结果应会加速开发炭疽芽孢杆菌检测方法以及了解芽孢衣生态作用的研究工作。
