Separation and structural analysis of lipoprotein in a lipopolysaccharide preparation from Porphyromonas gingivalis
Separation and structural analysis of lipoprotein in a lipopolysaccharide preparation from Porphyromonas gingivalis
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DOI：
10.1093/intimm/dxh146
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发表时间：
2004-10-01
影响因子：
4.4
通讯作者：
Ogawa, T
中科院分区：
文献类型：
--
作者：
Hashimoto, M;Asai, Y;Ogawa, T
牙周病细菌牙龈卟啉单胞菌(Pg-LPS)的脂多糖(LPS)制剂被认为需要Toll样受体(TLR)2而不是大肠杆菌LPS(EC-LPS)的受体TLR4来激活免疫细胞。然而，我们先前报道，牙龈假单胞菌的主要免疫刺激成分脂类A及其合成的类脂A通过TLR4依赖的途径激活细胞，而不是通过TLR2。在本研究中，PG-LPS中的一种脂蛋白(PG-LP)被证明是TLR2介导的细胞激活的主要成分。用疏水作用层析和制备电泳法分离Pg-Lp，经内肽测序鉴定为Pg-Lp，推测为牙龈假单胞菌基因组中编码的脂蛋白。用质谱仪对N-末端结构进行了表征，确定其为三酰化脂肽。PG-LP和EC-LPS均能有效地诱导人牙龈成纤维细胞产生IL-8。根据我们的结果，我们认为PG-LP是牙龈假单胞菌的一种强大的炎症因子。
Lipopolysaccharide (LPS) preparations from the periodontopathic bacterium Porphyromonas gingivalis (Pg-LPS) are thought to require Toll-like receptor (TLR)2 rather than TLR4, a receptor of Escherichia coli LPS (Ec-LPS), for activation of immune cells. However, we previously reported that P. gingivalis lipid A, an immunostimulatory principal component of LPS, and its synthetic counterpart activate cells through a TLR4-dependent pathway but not via TLR2. In the present study, a lipoprotein from Pg-LPS (Pg-LP) was shown to be a principal component for TLR2-mediated cell activation. Pg-LP was separated by hydrophobic interaction chromatography followed by preparative electrophoresis and identified by internal peptide sequencing as PG1828, a putative lipoprotein encoded in the P. gingivalis genome. The N-terminal structure was characterized as a triacylated lipopeptide using mass spectrometry. Pg-LP, as well as Ec-LPS, was potent in inducing IL-8 production in human gingival fibroblasts. From our results, we propose that Pg-LP is a powerful inflammatory factor of P. gingivalis.