Gibbs Karine A, Isaac Daniel D, Xu Jun, Hendrix Roger W, Silhavy Thomas J, Theriot Julie A
Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Mol Microbiol. 2004 Sep;53(6):1771-83. doi: 10.1111/j.1365-2958.2004.04242.x.
Advanced techniques for observing protein localization in live bacteria show that the distributions are dynamic. For technical reasons, most such techniques have not been applied to outer membrane proteins in Gram-negative bacteria. We have developed two novel live-cell imaging techniques to observe the surface distribution of LamB, an abundant integral outer membrane protein in Escherichia coli responsible for maltose uptake and for attachment of bacteriophage lambda. Using fluorescently labelled bacteriophage lambda tails, we quantitatively described the spatial distribution and dynamic movement of LamB in the outer membrane. LamB accumulated in spiral patterns. The distribution depended on cell length and changed rapidly. The majority of the protein diffused along spirals extending across the cell body. Tracking single particles, we found that there are two populations of LamB--one shows very restricted diffusion and the other shows greater mobility. The presence of two populations recalls the partitioning of eukaryotic membrane proteins between 'mobile' and 'immobile' populations. In this study, we have demonstrated that LamB moves along the bacterial surface and that these movements are restricted by an underlying dynamic spiral pattern.
用于观察活细菌中蛋白质定位的先进技术表明,蛋白质分布是动态的。由于技术原因,大多数此类技术尚未应用于革兰氏阴性菌的外膜蛋白。我们开发了两种新颖的活细胞成像技术,以观察LamB的表面分布,LamB是大肠杆菌中一种丰富的外膜整合蛋白,负责麦芽糖摄取和噬菌体λ的附着。使用荧光标记的噬菌体λ尾部,我们定量描述了LamB在外膜中的空间分布和动态运动。LamB以螺旋模式聚集。这种分布取决于细胞长度且变化迅速。大多数蛋白质沿着延伸穿过细胞体的螺旋扩散。追踪单个粒子时,我们发现有两类LamB——一类扩散非常受限,另一类具有更大的流动性。两类LamB的存在让人想起真核细胞膜蛋白在“可移动”和“不可移动”群体之间的分配。在本研究中,我们证明了LamB沿着细菌表面移动,并且这些移动受到潜在动态螺旋模式的限制。