Cleavage of host cytokeratin-6 by lysine-specific gingipain induces gingival inflammation in periodontitis patients.
Cleavage of host cytokeratin-6 by lysine-specific gingipain induces gingival inflammation in periodontitis patients.
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DOI：
10.1371/journal.pone.0117775
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Maruyama I
中科院分区：
文献类型：
--
作者：
Tancharoen S;Matsuyama T;Kawahara K;Tanaka K;Lee LJ;Machigashira M;Noguchi K;Ito T;Imamura T;Potempa J;Kikuchi K;Maruyama I
赖氨酸特异性牙龈菌蛋白酶（Lysine-specific gingipain，Kgp）是由牙龈卟啉单胞菌（Porphyromonas gingivalis，P. gingivalis）分泌的一种毒力因子。角蛋白中间丝维持牙龈上皮细胞的结构完整性，但被Kgp靶向产生新的细胞角蛋白6片段（K6 F）。我们研究了K6 F的释放及其对细胞因子分泌的诱导。K6 F存在于牙周病患者的龈沟液和牙龈卟啉酶处理的大鼠牙龈上皮细胞培养上清液中，通过基质辅助激光解吸/电离飞行时间质谱基于快速定量肽分析，使用BLOTCHIP测量。对牙龈组织进行K6 F免疫组化染色，采用免疫荧光染色和流式细胞术分析细胞角蛋白6蛋白的表达。免疫印迹法检测牙龈上皮细胞中MAPK的活化情况。ELISA法检测K6 F及其诱导的细胞因子释放。人牙龈成纤维细胞迁移进行了评估，使用Matrigel侵袭室测定。我们确定了K6 F，对应于人细胞角蛋白6的C-末端区域（氨基酸359-378），在牙周病患者的龈沟液和牙龈上皮细胞培养Kgp的上清液中。K6 F抗原在牙周病患者牙龈上皮中的分布从基底层到棘层。牙龈上皮细胞上的细胞角蛋白6可被Kgp降解，而不被Arg-牙龈卟啉单胞菌脂多糖、伴放线放线杆菌脂多糖降解。K6 F，而不是乱序K6 F肽，诱导人牙龈成纤维细胞迁移和分泌白细胞介素（IL）-6，IL-8和单核细胞趋化蛋白-1。K6 F的这些作用是通过激活p38 MAPK和Jun N-末端激酶介导的，而不是通过激活p42/44 MAPK或p-Akt介导的。Kgp将牙龈上皮细胞细胞角蛋白6降解为K6 F，K6 F在释放时诱导人牙龈成纤维细胞的侵袭和细胞因子分泌。因此，Kgp可能有助于牙周病的发展。
Lysine-specific gingipain (Kgp) is a virulence factor secreted from Porphyromonas gingivalis (P. gingivalis), a major etiological bacterium of periodontal disease. Keratin intermediate filaments maintain the structural integrity of gingival epithelial cells, but are targeted by Kgp to produce a novel cytokeratin 6 fragment (K6F). We investigated the release of K6F and its induction of cytokine secretion. K6F present in the gingival crevicular fluid of periodontal disease patients and in gingipain-treated rat gingival epithelial cell culture supernatants was measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer-based rapid quantitative peptide analysis using BLOTCHIP. K6F in gingival tissues was immunostained, and cytokeratin 6 protein was analyzed by immunofluorescence staining and flow cytometry. Activation of MAPK in gingival epithelial cells was evaluated by immunoblotting. ELISA was used to measure K6F and the cytokines release induced by K6F. Human gingival fibroblast migration was assessed using a Matrigel invasion chamber assay. We identified K6F, corresponding to the C-terminus region of human cytokeratin 6 (amino acids 359–378), in the gingival crevicular fluid of periodontal disease patients and in the supernatant from gingival epithelial cells cultured with Kgp. K6F antigen was distributed from the basal to the spinous epithelial layers in gingivae from periodontal disease patients. Cytokeratin 6 on gingival epithelial cells was degraded by Kgp, but not by Arg-gingipain, P. gingivalis lipopolysaccharide or Actinobacillus actinomycetemcomitans lipopolysaccharide. K6F, but not a scrambled K6F peptide, induced human gingival fibroblast migration and secretion of interleukin (IL)-6, IL-8 and monocyte chemoattractant protein-1. These effects of K6F were mediated by activation of p38 MAPK and Jun N-terminal kinase, but not p42/44 MAPK or p-Akt. Kgp degrades gingival epithelial cell cytokeratin 6 to K6F that, on release, induces invasion and cytokine secretion by human gingival fibroblasts. Thus, Kgp may contribute to the development of periodontal disease.
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影响因子：
3.7
作者：
Jünemann S;Prior K;Szczepanowski R;Harks I;Ehmke B;Goesmann A;Stoye J;Harmsen D
通讯作者：
Harmsen D
影响因子：
3.7
作者：
Dey M;Patra S;Su LY;Segall AM
通讯作者：
Segall AM
影响因子：
3.1
作者：
Dong, F;Su, H;Zhong, GM
通讯作者：
Zhong, GM
影响因子：
4.3
作者：
Hirose, M;Ishihara, K;Okuda, K
通讯作者：
Okuda, K
影响因子：
2.1
作者：
Fujimura, S;Hirai, K;Nakamura, T
通讯作者：
Nakamura, T