EFFECT OF FREE AND VESICLE-BOUND CYSTEINE PROTEINASES OF PORPHYROMONAS-GINGIVALIS ON PLASMA CLOT FORMATION - IMPLICATIONS FOR BLEEDING TENDENCY AT PERIODONTITIS SITES
EFFECT OF FREE AND VESICLE-BOUND CYSTEINE PROTEINASES OF PORPHYROMONAS-GINGIVALIS ON PLASMA CLOT FORMATION - IMPLICATIONS FOR BLEEDING TENDENCY AT PERIODONTITIS SITES
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DOI：
10.1128/iai.63.12.4877-4882.1995
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发表时间：
1995-12-01
影响因子：
3.1
通讯作者：
TRAVIS, J
中科院分区：
文献类型：
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作者：
IMAMURA, T;POTEMPA, J;TRAVIS, J
牙龈卟啉单胞菌感染与成人牙周炎密切相关，目前认为这种细菌的蛋白酶是重要的毒力因子。为了研究这些酶可能的病理功能，我们通过使用凝血酶时间（TT）测量来检查牙龈卟啉单胞菌的两种主要半胱氨酸蛋白酶（牙龈卟啉菌蛋白酶）的游离和囊泡结合形式对血浆凝块形成的影响。Lys-gingipain（gingipain-K）和Arg-gingipain（gingipain-R）均以剂量和时间依赖性方式延长血浆TT，并且这也在作为牙龈卟啉单胞菌蛋白酶的生物载体的囊泡中发现。血浆TT的囊泡的增加可以完全逆转治疗与非特异性半胱氨酸蛋白酶抑制剂，但只有部分选择性的化合物的牙龈蛋白酶-K或牙龈蛋白酶-R，预孵育的囊泡与牙龈蛋白酶-K-特异性抑制剂（z-FK-CK）减少血浆TT比牙龈蛋白酶-R-特异性抑制剂（亮抑酶肽），这表明在生理条件下，牙龈蛋白酶-K是更有效的纤维蛋白原破坏。每种纯化的酶也显著增加纤维蛋白原TT，牙龈卟啉菌蛋白酶-R比牙龈卟啉菌蛋白酶-K有效四倍。然而，在血浆中，牙龈菌蛋白酶-R是无效的，因为白蛋白的抑制作用。这些结果意味着，半胱氨酸蛋白酶，特别是牙龈蛋白酶-K，废除凝血功能的纤维蛋白原，因此，可能有助于出血的倾向，并在牙周炎部位感染牙龈卟啉单胞菌的持续炎症。
Infection by Porphyromonas gingivalis is strongly associated,vith adult periodontitis, with proteinases from this bacterium now considered to be important virulence factors. In order to investigate possible pathological functions of these enzymes, we examined the effect of both free and vesicle-bound forms of the two major cysteine proteinases (gingipains) of P. gingivalis on plasma clot formation by using thrombin time (TT) measurements. Both Lys-gingipain (gingipain-K) and Arg-gingipain (gingipain-R) prolonged plasma TT in a dose- and time-dependent manner, and this was also found with vesicles which are the biological carriers of P. gingivalis proteinases. The increase in plasma TT by vesicles could be completely reversed by treatment with nonspecific cysteine proteinase inhibitors but only partially by compounds selective for either gingipain-K or gingipain-R, Preincubation of vesicles with a gingipain-K-specific inhibitor (z-FK-ck) reduced plasma TT more than a gingipain-R-specific inhibitor (leupeptin), suggesting that under physiological conditions gingipain-K was more effective in fibrinogen destruction. Each purified enzyme also markedly increased fibrinogen TT, gingipain-R being fourfold more potent than gingipain-K. However, in plasma, gingipain-R was ineffective because of the inhibitory effect of albumin. These results imply that cysteine proteinases, especially gingipain-K, abrogate the clotting potential of fibrinogen and, therefore, may contribute to the bleeding tendency and to persistent inflammation in periodontitis sites infected with P. gingivalis.