Jiménez-Galisteo Guadalupe, Fusté Ester, Muñoz Elisa, Vinuesa Teresa, Villa Tom G, Benz Roland, Domínguez Angel, Viñas Miguel
Department of Pathology and Experimental Therapeutics, Medical School, IDIBELL-University of Barcelona.
Department of Public Health, Mental Health and Perinatal Nursing, IDIBELL-University of Barcelona.
J Gen Appl Microbiol. 2017 Nov 17;63(5):266-273. doi: 10.2323/jgam.2017.01.001. Epub 2017 Aug 23.
Gordonia jacobaea is a bacterium belonging to the mycolata group characterized by its ability to produce carotenoids. Mycolic acids in the cell wall contribute to reducing the permeability of their envelopes requiring the presence of channel-forming proteins to allow the exchange of hydrophilic molecules with the surrounding medium. Identification and purification of the channel-forming proteins was accomplished by SDS-PAGE, Mass spectrometry and Mass peptide fingerprinting and the channel-forming activity was studied by reconstitution in lipid bilayers. Here, we describe for the first time the presence of a cell-wall protein from G. jacobaea with channel-forming activity. Our results suggest that this protein bears a low similarity to other hypothetical proteins from the genus Gordonia of uncharacterized functions. The channel has an average single-channel conductance of 800 pS in 1 M KCl, is moderately anion-selective, and does not show any voltage dependence for voltages between +100 and -100 mV. The channel characteristics suggest that this protein could be of relevance in the import and export of negatively charged molecules across the cell wall. This could contribute to design treatments for mycobacterial infections, as well as being of interest in biotechnology applications.
黄顶菊戈登氏菌是一种属于分枝杆菌类的细菌,其特点是能够产生类胡萝卜素。细胞壁中的分枝菌酸有助于降低其包膜的通透性,这就需要存在形成通道的蛋白质来允许亲水分子与周围介质进行交换。通过SDS-PAGE、质谱分析和质谱肽指纹图谱完成了对形成通道蛋白质的鉴定和纯化,并通过在脂质双层中重组来研究通道形成活性。在此,我们首次描述了黄顶菊戈登氏菌中一种具有通道形成活性的细胞壁蛋白的存在。我们的结果表明,该蛋白与戈登氏菌属其他功能未明的假设蛋白相似度较低。该通道在1 M KCl中的平均单通道电导为800 pS,具有中等程度的阴离子选择性,并且在+100至-100 mV的电压范围内未显示出任何电压依赖性。通道特性表明,该蛋白可能与带负电荷分子跨细胞壁的进出有关。这可能有助于设计针对分枝杆菌感染的治疗方法,以及在生物技术应用中具有重要意义。