Aggregation of human platelets by gingipain-R from Porphyromonas gingivalis cells and membrane vesicles

Platelets
K PhamE J Whitaker

Abstract

The hypothesis that there is an association between periodontitis and cardiovascular disease suggests new lines of research on the mechanism whereby oral bacteria might exert systemic effects. This study was conducted to ascertain and quantitate the effect of Porphyromonas gingivalis on human platelets in vitro. A second related objective was to purify and identify the aggregating vector. Aggregation was measured by platelet turbidometry and gingipain-R was purified from P. gingivalis membrane vesicles by Sepharose 2B and hydroxyapatite chromatography. The in vitro aggregation of platelets requires that at least 1.0 x 10(4) cells be stirred with 1.35 x 10(8) platelets. The specific activity is substantially increased in the membrane vesicles that are shed by this bacterium. Aggregability was due to gingipain-R activity, a potent cysteine protease that was found to be highly concentrated in the membrane vesicle fraction. The enzyme was purified 18-fold in high yield from the membrane vesicles, and consists of two noncovalently linked proteins that migrate at 49 and 44 kDa on SDS-PAGE. Aggregation of platelets by gingipain-R was shown to be dose-dependent, and inhibited by leupeptin and arginine, but not by anti-thrombin III. Thi...Continue Reading

Citations

Jan 19, 2011·Journal of Cardiovascular Pharmacology·Jing XiongMuthuvel Jayachandran
Feb 8, 2006·Journal of Periodontology·A ContrerasJ E Botero
Jan 1, 2009·Journal of Oral Microbiology·Steven W Kerrigan, Dermot Cox
Jun 18, 2010·Canadian Journal of Physiology and Pharmacology·Archibald McNicol, Sara J Israels
Nov 23, 2006·Journal of Thrombosis and Haemostasis : JTH·P JäremoA Richter
Apr 3, 2018·Journal of Clinical Periodontology·Yvonne Jockel-SchneiderMartin Eigenthaler
Apr 23, 2005·Arteriosclerosis, Thrombosis, and Vascular Biology·Nadine BrodalaTimothy C Nichols
May 21, 2019·Frontiers in Neurology·Priscila Cunha NascimentoRafael Rodrigues Lima
Sep 20, 2020·Molecular Oral Microbiology·William A ChenDanilo S Boskovic
Oct 9, 2021·Seminars in Thrombosis and Hemostasis·Martin J Page, Etheresia Pretorius

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