Thein Marcus, Bunikis Ignas, Denker Katrin, Larsson Christer, Cutler Sally, Drancourt Michel, Schwan Tom G, Mentele Reinhard, Lottspeich Friedrich, Bergström Sven, Benz Roland
Department of Biotechnology, Biocenter, University of Würzburg, Am Hubland, D-97074 Würzburg, Germany.
J Bacteriol. 2008 Nov;190(21):7035-42. doi: 10.1128/JB.00818-08. Epub 2008 Aug 29.
Relapsing fever is a worldwide, endemic disease caused by several spirochetal species belonging to the genus Borrelia. During the recurring fever peaks, borreliae proliferate remarkably quickly compared to the slow dissemination of Lyme disease Borrelia and therefore require efficient nutrient uptake from the blood of their hosts. This study describes the identification and characterization of the first relapsing fever porin, which is present in the outer membranes of B. duttonii, B. hermsii, B. recurrentis, and B. turicatae. The pore-forming protein was purified by hydroxyapatite chromatography and designated Oms38, for outer membrane-spanning protein of 38 kDa. Biophysical characterization of Oms38 was done by using the black lipid bilayer method, demonstrating that Oms38 forms small, water-filled channels of 80 pS in 1 M KCl that did not exhibit voltage-dependent closure. The Oms38 channel is slightly selective for anions and shows a ratio of permeability for cations over anions of 0.41 in KCl. Analysis of the deduced amino acid sequences demonstrated that Oms38 contains an N-terminal signal sequence which is processed under in vivo conditions. Oms38 is highly conserved within the four studied relapsing fever species, sharing an overall amino acid identity of 58% and with a strong indication for the presence of amphipathic beta-sheets.
回归热是一种由几种属于疏螺旋体属的螺旋体引起的全球性地方病。在反复发热高峰期,与莱姆病疏螺旋体的缓慢传播相比,疏螺旋体增殖非常迅速,因此需要从宿主血液中高效摄取营养物质。本研究描述了首个回归热孔蛋白的鉴定和特征,该蛋白存在于达顿疏螺旋体、赫氏疏螺旋体、回归热疏螺旋体和图莱里疏螺旋体的外膜中。通过羟基磷灰石色谱法纯化了形成孔道的蛋白,并将其命名为Oms38,即38 kDa的外膜跨膜蛋白。采用黑脂质双层法对Oms38进行了生物物理特性鉴定,结果表明Oms38在1 M KCl中形成了80 pS的小的充满水的通道,且不表现出电压依赖性关闭。Oms38通道对阴离子有轻微选择性,在KCl中阳离子与阴离子的渗透率之比为0.41。对推导的氨基酸序列分析表明,Oms38含有一个N端信号序列,该序列在体内条件下会被加工。Oms38在所研究的四种回归热物种中高度保守,总体氨基酸同一性为58%,且强烈提示存在两亲性β折叠。