EXTRACELLULAR VESICLE-ASSOCIATED AND SOLUBLE TRYPSIN-LIKE-ENZYME FRACTIONS OF PORPHYROMONAS-GINGIVALIS W50

被引：17

作者：

SMALLEY, JW

BIRSS, AJ

机构：

[1] Department of Clinical Dental Sciences, University of Liverpool, Liverpool

来源：

ORAL MICROBIOLOGY AND IMMUNOLOGY | 1991年 / 6卷 / 04期

关键词：

PORPHYROMONAS-GINGIVALIS; PROTEASE; EXTRACELLULAR VESICLE;

D O I：

10.1111/j.1399-302X.1991.tb00478.x

中图分类号：

R78 [口腔科学];

学科分类号：

1003 ;

摘要：

Soluble vesicle-associated trypsin-like enzyme fractions (VSF) were prepared by sonication from extracellular vesicles (ECV) from strains W50 and W50/BE1. High-(H), intermediate-(I) and low-(L) molecular-weight VSF enzyme subfractions were identified by non-dissociative gel filtration chromatography with Mr 160, 95 and 60 kDa respectively. The chromatographic profiles of W50 VSF from 48-h and 72-h cultures were identical. W50/BE1 VSF displayed a higher ratio of the 160 to 60 kDa components. This ratio was reduced in VSF from 72-h cultures. Extracellular soluble protein (EP) trypsin profiles were similar to their respective VSF, but the 60 kDa component predominated. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed a loss of soluble extracellular polypeptides with culture age. A polyclonal antiserum to EP subfraction L reacted in immunoblots with a 50 kDa peptide of subfraction L of W50. Whole EP and its subfraction H displayed a 50 kDa immunoreactive peptide but no peptides of higher molecular weight. This antiserum reacted with a similar sized peptide, and with lower-molecular-weight components in whole ECV. Gelatin substrate zymography of whole EP following non-reducing SDS-PAGE revealed a major 80 kDa protease that increased with culture age. Minor protease bands of 70 and 50 kDa were also detected.

引用

页码：202 / 208

页数：7

共 24 条

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