Comparative properties of envelope-associated arginine-gingipains and lysine-gingipain of Porphyromonas gingivalis
Comparative properties of envelope-associated arginine-gingipains and lysine-gingipain of Porphyromonas gingivalis
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DOI：
10.1016/s0378-1097(98)00169-4
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发表时间：
1998-06-15
影响因子：
2.1
通讯作者：
Nakamura, T
中科院分区：
文献类型：
--
作者：
Fujimura, S;Hirai, K;Nakamura, T
通过离子交换层析、亲和层析和等电聚焦等步骤，从经洗涤剂增溶的牙龈卟啉单胞菌的包膜材料中分离纯化出两种精氨酸特异性蛋白水解酶(RGP-A、RGP-B)和一种赖氨酸特异性蛋白水解酶(Lys-gigiain[KGP])。每种纯化的酶在SDS-PAGE上均显示一条单一的染色条带。这三种酶通常被硫醇、二硫苏糖醇和半胱氨酸等还原试剂激活。甘氨酰甘氨酸可显著激活RGP-B，EDTA可显著激活KGP。对羟基苯甲酸汞、对甲苯磺酰-L-赖氨酸氯甲基酮、N-乙基马来酰亚胺、乙二胺四乙酸二乙酯、亮肽素、L-trans-epoxy-succinylleucylamido-(4-guanidino)butane和止痛剂均能抑制Ph RGP-A和B的表达。对羟基苯甲酸汞、N-乙基马来酰亚胺和TLCK对KGP有抑制作用。RGP-A和RGP-B的相对分子质量相同(43 KDa)，KGP的相对分子质量为48 kDa。RGPS和KGP对合成显色底物的水解活性仅限于P-1位含有精氨酸和赖氨酸的化合物。当这三种酶分别作用于免疫球蛋白时，在每个反应产物中都产生了两个新的34 kDa和15 kDa的片段。RGP-B几乎完全消化了Hb，但RGP-A和KGP处理不能引起Hb的实质性降解。(C)1998年，爱思唯尔科学公司出版。保留所有权利。
Two arginine specific proteinases (Arg-gingipain [RGP-A, RGP-B]) and a lysine specific proteinase (Lys-gingipain [KGP]) were purified from materials of the envelope of Porphyromonas gingivalis solubilized by a detergent by the sequential procedures of ion-exchange chromatography, affinity chromatography, and isoelectric focusing. Each purified enzyme showed a single stained band on SDS-PAGE. The three enzymes were commonly activated by reducing reagents such as mercaptoethanol, dithiothreitol and cysteine. RGP-B was activated markedly by glycyl-glycine and KGP was activated significantly by EDTA. Po th RGP-A and RGP-B were inhibited by p-hydroxymercuribenzoate, tosyl-L-lysine chloromethyl ketone (TLCK), N-ethylmaleimide, EDTA, leupeptin, L-trans-epoxy-succinylleucylamido-(4-guanidino)butane and antipain. KGP was inhibited by p-hydroxymercuribenzoate, N-ethylmaleimide and TLCK. The molecular masses of RGP-A and RGP-B were the same (43 kDa), and that of KGP was 48 kDa. The hydrolytic activities of RGPs and KGP to chromogenic synthetic substrates were limited to the compounds with arginine and lysine in the P-1 positions, respectively. When IgG was treated with the three enzymes separately, it was demonstrated that two new fragments of 34 kDa and 15 kDa were generated in each reaction product. Hemoglobin was almost exhaustively digested by RGP-B, but substantial degradation could not be induced by RGP-A or KGP treatment. (C) 1998 Published by Elsevier Science B.V. All rights reserved.