[牙龈卟啉单胞菌W50细胞外囊泡中牙龈毒素的研究：纯化及细胞毒性特征分析]

[A study of gingivain from extracellular vesicles of Porphyromonas gingivalis W50: purification and cytotoxic characterization].

作者信息

Ni L, Shi J, Liu Z

机构信息

Stomatological College, Fourth Military Medical University, Xi'an.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 1996 Mar;31(2):95-8.

PMID:9387541

Abstract

Gingivain was purified from extracellular vesicles of Porphyromonas gingivalis W50 with high performance liquid chromatography (HPLC) and its cytotoxic characterization on L929 cell line was examined. The result showed that gingivain might belong to systeine proteinases. The molecular weight of gingivain was about 27,800 unit by HPLC in which there might be one or two peptides being 10,900 unit, the enzyme was activated by Ca2+ and thiol-containing agents such as dithiothycitol and inhibited by Na-4-tosyl-L-lysine chioromethyl and EDTA. The maximum activity of the enzyme were found at pH 7.5, purified gingivain was toxic to L929 cell line at the content of 10 micrograms/ml. The study indicates that gingivain carried by vesicles might be more important in bacterial toxic activities.

摘要

利用高效液相色谱法（HPLC）从牙龈卟啉单胞菌W50的细胞外囊泡中纯化牙龈蛋白酶，并检测其对L929细胞系的细胞毒性特征。结果表明，牙龈蛋白酶可能属于半胱氨酸蛋白酶。通过HPLC测定，牙龈蛋白酶的分子量约为27,800道尔顿，其中可能有一或两条肽链为10,900道尔顿，该酶被Ca2+和含硫醇试剂（如二硫苏糖醇）激活，并被对甲苯磺酰-L-赖氨酸氯甲基酮和乙二胺四乙酸（EDTA）抑制。该酶的最大活性出现在pH 7.5时，纯化后的牙龈蛋白酶在浓度为10微克/毫升时对L929细胞系有毒性。研究表明，由囊泡携带的牙龈蛋白酶在细菌毒性活动中可能更为重要。