Attenuation of the virulence of Porphyromonas gingivalis by using a specific synthetic kgp protease inhibitor 2
Attenuation of the virulence of Porphyromonas gingivalis by using a specific synthetic kgp protease inhibitor 2
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DOI：
10.1128/iai.70.12.6968-6975.2002
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发表时间：
2002-12-01
影响因子：
3.1
通讯作者：
Samuelsson, B
中科院分区：
文献类型：
--
作者：
Curtis, MA;Opoku, JA;Samuelsson, B
牙龈卟啉单胞菌的精氨酸和赖氨酸牙龈卟啉蛋白酶是牙周病中重要的毒力决定因素，可能对应于基于免疫或药物的治疗策略的目标。在这项研究中，我们的目的是确定这些酶代表最有前途的分子靶蛋白酶的治疗，并检查所得化合物在小鼠毒力测定的有效性。通过使用牙龈卟啉单胞菌W50制备在rgpA和rgpB（编码Arg-牙龈卟啉菌蛋白酶）和kgp（编码Lys-牙龈卟啉菌蛋白酶）中具有突变的同基因突变体以及在rgpA和rgpB中具有突变的双突变体。这些突变体的毒力表明，Kgp是一个有前途的药物靶点。组合化学用于确定Kgp的最佳底物，并根据此信息设计和合成具有纳摩尔K-i的特异性缓慢可逆抑制剂。牙龈卟啉单胞菌W50在该化合物存在下的生长类似于kgp同基因突变体的表型;在两种情况下，细菌菌落均未能在血琼脂上形成色素，并且在含有白蛋白作为唯一蛋白质来源的限定培养基中仅获得较差的生长。此外，用Kgp抑制剂预处理野生型微生物导致小鼠试验中毒力显著降低。这些数据强调了Kgp是牙龈卟啉单胞菌营养和毒力的重要因素的结论，并且该酶的抑制剂可能具有控制牙龈卟啉单胞菌感染的治疗潜力。蛋白酶抑制剂可能是一类潜在的新型抗菌剂，与其他细菌病原体的控制有关。
The Arg- and Lys-gingipains of Porphyromonas gingivalis are important virulence determinants in periodontal disease and may correspond to targets for immune- or drug-based treatment strategies. In this investigation we aimed to determine which of these enzymes represents the most promising molecular target for protease inhibitor-based therapy and to examine the effectiveness of the resultant compound in a murine virulence assay. Isogenic mutants with mutations in rgpA and rgpB (encoding Arg-gingipains) and in kgp (encoding Lys-gingipain) and a double mutant with mutations in rgpA and rgpB were prepared by using P. gingivalis W50. The virulence of these mutants indicated that Kgp is a promising drug target. Combinatorial chemistry was used to define the optimal substrate of Kgp, and from this information a specific slowly reversible inhibitor with a nanomolar K-i was designed and synthesized. Growth of P. gingivalis W50 in the presence of this compound resembled the phenotype of the kgp isogenic mutant; in both instances bacterial colonies failed to form pigment on blood agar, and only poor growth was obtained in a defined medium containing albumin as the sole protein source. Furthermore, pretreatment of the wild-type organism with the Kgp inhibitor led to a significant reduction in virulence in the murine assay. These data emphasize the conclusion that Kgp is an important factor for both nutrition and virulence of P. gingivalis and that inhibitors of this enzyme may have therapeutic potential for the control of P. gingivalis infections. Protease inhibitors may be a potentially novel class of antimicrobial agents with relevance to the control of other bacterial pathogens.