Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.
Mol Oral Microbiol. 2013 Jun;28(3):181-91. doi: 10.1111/omi.12013. Epub 2012 Dec 17.
Interstrain differences in antigenic surface proteins may reflect immunological pressure or differences in receptor specificity of the antigen. Treponema denticola exhibits considerable interstrain variability in its major surface protein (Msp), but no studies have addressed this issue in dentilisin (CTLP), a surface protease complex that has a significant role in T. denticola-host interactions in periodontal disease. Furthermore, the genome annotation of the prcB-prcA-prtP operon encoding dentilisin contains apparent errors and lacks a deduced PrtP amino acid sequence. To address these issues we analysed the protease operon from diverse T. denticola strains, as well as clones of the ATCC 35405 Type strain from which the genome sequence and original GenBank prtP sequence were derived. 6xHis-tagging of the PrtP C-terminus in ATCC 35405 demonstrated absence of the 'authentic frameshift' in PrtP reported in the genome databases. We propose that T. denticola genome annotations be updated to reflect this new information. PrcB and the PrtP N-terminal region that includes the catalytic domain were highly conserved in common laboratory strains and clinical isolates of T. denticola. Dentilisin proteolytic activity varied considerably between strains. Antibodies against PrcB, PrcA and PrtP from the type strain recognized these proteins in most T. denticola strains. PrtP varied up to 20% over the C-terminal 270 residues between strains. The PrtP C-terminal eight-residues (DWFYVEYP) was present in all strains, with two strains containing an additional Y-residue preceding the stop codon. Such conserved PrtP domains may be required for interactions with PrcA and PrcB, or for substrate interactions.
不同菌株间抗原表面蛋白的差异可能反映了免疫压力或抗原受体特异性的差异。齿垢密螺旋体在其主要表面蛋白(Msp)中表现出相当大的菌株间变异性,但尚无研究探讨牙蛋白酶(CTLP)在牙周病中与宿主相互作用方面具有重要作用的表面蛋白酶复合物中的这一问题。此外,编码牙蛋白酶的 prcB-prcA-prtP 操纵子的基因组注释存在明显错误,并且缺乏推导的 PrtP 氨基酸序列。为了解决这些问题,我们分析了来自不同齿垢密螺旋体菌株的蛋白酶操纵子,以及源自基因组序列和原始 GenBank prtP 序列的 ATCC 35405 型菌株的克隆。在 ATCC 35405 中对 PrtP C 末端进行 6xHis 标记表明,在基因组数据库中报道的 PrtP 中不存在“真实移码”。我们建议更新齿垢密螺旋体基因组注释以反映这一新信息。PrcB 和包括催化结构域的 PrtP N 末端在常见的实验室菌株和临床分离株中高度保守。牙蛋白酶的蛋白水解活性在菌株间差异很大。来自型菌株的针对 PrcB、PrcA 和 PrtP 的抗体在大多数齿垢密螺旋体菌株中识别这些蛋白。PrtP 在菌株间的 C 末端 270 个残基之间差异高达 20%。PrtP 的 C 末端 8 个残基(DWFYVEYP)存在于所有菌株中,有两个菌株在终止密码子之前含有另外一个 Y 残基。这些保守的 PrtP 结构域可能是与 PrcA 和 PrcB 相互作用所必需的,或者是与底物相互作用所必需的。