Por secretion system-dependent secretion and glycosylation of Porphyromonas gingivalis hemin-binding protein 35.
Por secretion system-dependent secretion and glycosylation of Porphyromonas gingivalis hemin-binding protein 35.
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DOI：
10.1371/journal.pone.0021372
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Nakayama K
中科院分区：
文献类型：
--
作者：
Shoji M;Sato K;Yukitake H;Kondo Y;Narita Y;Kadowaki T;Naito M;Nakayama K
牙龈卟啉单胞菌（Porphyromonas gingivalis）是严重牙周病和难治性根尖周炎的主要病原菌。我们最近发现牙龈卟啉单胞菌具有一种新的分泌系统，称为Por分泌系统（PorSS），其负责分泌主要的胞外蛋白酶，Arg-gingipain（Rgps）和Lys-gingipain。这些蛋白酶在其C-末端含有保守的C-末端结构域（CTD）。血红素结合蛋白35（HBP35）是牙龈卟啉单胞菌的外膜蛋白之一，有助于其血红素利用，也包含CTD，表明HBP35通过PorSS易位到细胞表面。在这项研究中，免疫印迹分析的牙龈卟啉单胞菌突变体的缺陷，在PorSS或阴离子多糖-脂多糖（A-LPS）的生物合成显示，HBP35易位到细胞表面通过PorSS和A-LPS的糖基化。从用GFP-CTD [HBP35]绿色荧光蛋白融合物的缺失分析，发现CTD [HBP35]的C-末端22个氨基酸残基是细胞表面易位和糖基化所需的。GFP-CTD融合研究还显示CPG70、肽基精氨酸脱亚胺酶、P27和RgpB的CTD在PorSS依赖的易位和糖基化中起作用。然而，CTD区肽的糖基化HBP35蛋白的样品中没有发现肽图谱指纹分析，和CTD区肽的抗体不与糖基化HBP35蛋白反应。这些结果表明，CTD区域的功能作为识别信号的PorSS和CTD蛋白的糖基化后发生的CTD区域的去除。本研究用家兔制备抗细菌蛋白的抗血清。
The anaerobic Gram-negative bacterium Porphyromonas gingivalis is a major pathogen in severe forms of periodontal disease and refractory periapical perodontitis. We have recently found that P. gingivalis has a novel secretion system named the Por secretion system (PorSS), which is responsible for secretion of major extracellular proteinases, Arg-gingipains (Rgps) and Lys-gingipain. These proteinases contain conserved C-terminal domains (CTDs) in their C-termini. Hemin-binding protein 35 (HBP35), which is one of the outer membrane proteins of P. gingivalis and contributes to its haem utilization, also contains a CTD, suggesting that HBP35 is translocated to the cell surface via the PorSS. In this study, immunoblot analysis of P. gingivalis mutants deficient in the PorSS or in the biosynthesis of anionic polysaccharide-lipopolysaccharide (A-LPS) revealed that HBP35 is translocated to the cell surface via the PorSS and is glycosylated with A-LPS. From deletion analysis with a GFP-CTD[HBP35] green fluorescent protein fusion, the C-terminal 22 amino acid residues of CTD[HBP35] were found to be required for cell surface translocation and glycosylation. The GFP-CTD fusion study also revealed that the CTDs of CPG70, peptidylarginine deiminase, P27 and RgpB play roles in PorSS-dependent translocation and glycosylation. However, CTD-region peptides were not found in samples of glycosylated HBP35 protein by peptide map fingerprinting analysis, and antibodies against CTD-regions peptides did not react with glycosylated HBP35 protein. These results suggest both that the CTD region functions as a recognition signal for the PorSS and that glycosylation of CTD proteins occurs after removal of the CTD region. Rabbits were used for making antisera against bacterial proteins in this study.
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DOI：
10.1016/s0306-3623(96)00366-7
发表时间：
1997-05-01
期刊：
GENERAL PHARMACOLOGY
影响因子：
--
作者：
Saito, S;Hiratsuka, K;Abiko, Y
通讯作者：
Abiko, Y
影响因子：
3.1
作者：
Kondo, Yoshio;Ohara, Naoya;Nakayama, Koji
通讯作者：
Nakayama, Koji
影响因子：
2.8
作者：
Sato, Keiko;Kido, Nobuo;Yoshimura, Fuminobu
通讯作者：
Yoshimura, Fuminobu
影响因子：
3.6
作者：
Paramonov, N;Rangarajan, M;Curtis, MA
通讯作者：
Curtis, MA
DOI：
10.1073/pnas.0500044102
发表时间：
2005-02-22
影响因子：
11.1
作者：
Feldman, MF;Wacker, M;Aebi, M
通讯作者：
Aebi, M