Suppression of pathogenicity of Porphyromonas gingivalis by newly developed gingipain inhibitors
Suppression of pathogenicity of Porphyromonas gingivalis by newly developed gingipain inhibitors
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DOI：
10.1124/mol.104.004366
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发表时间：
2004-12-01
影响因子：
3.6
通讯作者：
Yamamoto, K
中科院分区：
文献类型：
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作者：
Kadowaki, T;Baba, A;Yamamoto, K
Arg-gingipain （Rgp）和Lys-gingipain （Kgp）是由牙龈卟啉单胞菌产生的半胱氨酸蛋白酶，是牙周病的主要致病细菌。在这里，我们展示了一系列能够抑制Rgp或Kgp的小肽类似物，它们是根据每个酶对人唾液组蛋白的裂解位点特异性合成的。其中，carbobenzoxy-Lys-Arg-CO-Lys-N(CH3)(2) （KYT-1）和carbobenzoxy-Glu(NHN(CH3) Ph)- lys - co - NHCH2Ph （KYT-36）分别是Rgp和Kgp最有效的抑制剂，其K-i值为10(-11)~ 10(-10)M级。这两种抑制剂对哺乳动物蛋白酶如胰蛋白酶、组织蛋白酶B、L和h均表现出轻微或无抑制作用。所有由牙龈假单胞菌培养上清诱导的毒力测试，包括各种宿主蛋白的降解，如人类I型胶原蛋白、免疫球蛋白、纤维连接蛋白和纤维蛋白原，破坏多形核白细胞的杀菌活性，增强血管通透性。被两种抑制剂的联合作用强烈抑制。细菌在牙周袋中生长和存活所必需的功能，如氨基酸的共聚集和获取，也被两种抑制剂的联合作用强烈抑制。单次使用KYT-1可强烈抑制人成纤维细胞粘附和活力的破坏以及机体的血凝。这些结果表明，新开发的KYT-1和KYT-36都应该在这类重要酶的研究中提供更广泛的应用，并促进牙周病新方法的发展。
Arg-gingipain (Rgp) and Lys-gingipain (Kgp) are cysteine proteinases produced by Porphyromonas gingivalis, a major etiological bacterium of periodontal diseases. Here we show a series of small peptide analogs able to inhibit either Rgp or Kgp, which are synthesized on the basis of the cleavage site specificity of human salivary histatins by each enzyme. Among this series of compounds, carbobenzoxy-Lys-Arg-CO-Lys-N( CH3)(2) (KYT-1) and carbobenzoxy-Glu(NHN(CH3) Ph)-Lys-CO- NHCH2Ph (KYT-36) were found to be the most potent inhibitors of Rgp and Kgp, respectively, with K-i values of 10(-11) to 10(-10) M order. Both inhibitors exhibited slight or no inhibition on mammalian proteinases such as trypsin and cathepsins B, L, and H. All of the virulence induced by the culture supernatant of P. gingivalis tested, including the degradation of various host proteins such as human type I collagen, immunoglobulins, fibronectin, and fibrinogen, disruption of the bactericidal activity of polymorphonuclear leukocytes, and enhancement of the vascular permeability, were strongly inhibited by a combined action of both inhibitors. The functions essential for the bacterium to grow and survive in the periodontal pocket, such as coaggregation and acquisition of amino acids, were also strongly inhibited by the combined action of both inhibitors. The disruption of the adhesion and viability of human fibroblasts and hemagglutination by the organism were strongly suppressed by a single use of KYT-1. These results thus indicate that the newly developed KYT-1 and KYT-36 both should provide a broader application in studies of this important class of enzymes and facilitate the development of new approaches to periodontal diseases.