Inhibitory effect of human α_2-plasmin inhibitor and it's intensified compound by S-nitrosylation on the activity of arginine-specific cysteine proteinase from Porphyromonas gingivalis.

人α_2-纤溶酶抑制剂及其S-亚硝基化强化化合物对牙龈卟啉单胞菌精氨酸特异性半胱氨酸蛋白酶活性的抑制作用。

• 批准号: 13671931
• 负责人: KAMINISHI Hidenori
• 金额: $ 1.47万
• 依托单位: Fukuoka Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671931/
• 关键词: P. gingivalis; gingipain; α_2-plasmin inhibitor; S-nitrosylation; α2アンチプラスミン; α1プロテアーゼインヒビター; ベジクル; vesicle; α-2アンチプラスミン; Porphyromonas gingivalis

We investigated the effect of human α_2-plasmin inhibitor (α_2PI) and S-nitrosylated α_2-plasmin inhibitor (S-NO-α_2PI) on the activity of arginine-specific cysteine proteinase (gingipain, RGP) from Porphyromonas gingivalis, an aetiological bacterium of adult periodontitis, with the use of fluorogenic substrates. We discovered that human α_2PI as well as S-NO-α_2PI play a inhibitory effect to the RGP in a dose-dependent manner. The α_2PI, one of the members of serpin family, is well known as a specific inhibitor of plasmin, a typical serine proteinase. However, few studies have been reported concerning the inhibitory potency of humoral factors, such as α_1-protease inhibitor or α_2-macroglobulin, against cysteine proteinases especially RGP. In this study, we found a novel function of α_2PI as a potent inhibitor of RGP. The inhibitory potencies of the S-nitrosylated derivative against both human plasmin and RGP were slightly higher than those of the intact α_2PI. These results strongly suggest that S-NO-α_2PI is a useful tool for preventing the virulence of P. gingivalis.

本实验采用荧光底物法研究了人α_2纤溶酶抑制剂（α_2PI）和S-亚硝基化α_2纤溶酶抑制剂（S-NO-α_2PI）对牙龈卟啉单胞菌（Porphyromonas gingivalis）的牙龈蛋白酶（gingipain，RGP）活性的影响。我们发现人α_2PI和S-NO-α_2PI对RGP有抑制作用，并呈剂量依赖性。α_2PI是丝氨酸蛋白酶抑制剂家族的成员之一，是纤溶酶的特异性抑制剂。然而，关于体液因子如α_1-蛋白酶抑制剂或α_2-巨球蛋白对半胱氨酸蛋白酶特别是RGP的抑制作用的研究报道很少。本研究发现α_2PI作为RGP的有效抑制剂具有新的功能。S-亚硝基化衍生物对人纤溶酶和RGP的抑制作用略高于完整的α_2PI。提示S-NO-α_2PI可作为抑制牙龈卟啉单胞菌毒力的有效工具。

项目成果

上西秀則 ほか: "真菌の遺伝学"日本臨牀. 61・3. 785-792 (2003)

Hidenori Uenishi 等：“真菌遗传学”Nippon Rinban 61・3（2003）。

Cho, T., M.Sudoh, H, Kaminishi et al.: "Isolation and sequencing of the Candida albicans MS13, a putative novel member of the HSP70 family"Yeast. 20. 149-156 (2003)

Cho, T., M.Sudoh, H, Kaminishi 等人：“白色念珠菌 MS13（HSP70 家族的假定新成员）酵母的分离和测序”。

上西秀則, 小倉理恵子 ほか: "α_2プラスミンインヒビターおよびそのニトロソ化物はPorphyromonas gingivalisプロテアーゼの活性を阻害する"医学と生物学. 148. 27-31 (2004)

Hidenori Uenishi、Rieko Ogura 等人：“α_2 纤溶酶抑制剂及其亚硝酸盐抑制牙龈卟啉单胞菌蛋白酶的活性”医学和生物学 148. 27-31 (2004)。

Inoue Y, Fukushima T., et al.: "Antibacterial characteristics of newly developed amphiphilic lipids and DNA-lipid complexes against bacteria."J.Biomed.Mater.Res.. 65A. 203-208 (2003)

Inoue Y、Fukushima T. 等人：“新开发的两亲性脂质和 DNA-脂质复合物对细菌的抗菌特性。”J.Biomed.Mater.Res. 65A。

上西秀則, 長 環, 小倉理恵子: "真菌の遺伝学"日本臨牀. 61. 785-792 (2003)

Hidenori Uenishi、Tamaki Naga、Rieko Ogura：“真菌遗传学”Nippon Rinburi 61. 785-792 (2003)。