Characterization of Porphyromonas gingivalis sialidase and disruption of its role in host-pathogen interactions
Characterization of Porphyromonas gingivalis sialidase and disruption of its role in host-pathogen interactions
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DOI：
10.1099/mic.0.000851
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发表时间：
2019-11-01
期刊：
影响因子：
2.8
通讯作者：
Stafford, Graham
中科院分区：
文献类型：
--
作者：
Frey, Andrew M.;Satur, Marianne J.;Stafford, Graham
牙周炎发病和进展的关键是口腔细菌与宿主之间的复杂关系。与严重牙周炎最相关的微生物是红色复合体的牙周病原体：Tannerella denticola和Porphyromonas gingivalis。这些生物表达唾液酸酶，其从宿主糖蛋白切割唾液酸，并通过各种机制促成疾病。在这里，我们表达并纯化重组牙龈卟啉单胞菌唾液酸酶SiaPG（PG_0352），并表征其对多种底物的活性，包括宿主唾液酸糖蛋白，并突出显示不能切割二乙酰化唾液酸-这一现象被来自T.艾西娅事实上，SiaPG需要NanS来最大限度地从重度O-乙酰化的底物（如牛唾液粘蛋白）中收获唾液酸，这暗示了口腔微生物群的这些成员在从宿主来源释放唾液酸方面的种间合作的可能性。使用市售化疗剂扎那米韦抑制SiaPG和牙龈卟啉单胞菌的活性，表明其作为毒力抑制剂的潜力，其还抑制唾液酸从粘蛋白释放，并且能够抑制牙龈卟啉单胞菌在口腔糖蛋白来源上的生物膜形成。扎那米韦还在单种和多种感染实验中抑制牙龈卟啉单胞菌和其他牙周病原体对口腔上皮细胞的附着和侵袭，表明其具有抑制混合微生物群落的宿主-病原体相互作用的潜力。这项研究拓宽了我们对细菌唾液酸酶在毒力中的多种作用的理解，并表明用化疗药物抑制唾液酸酶可能是牙周炎治疗的一个有前途的策略。
Key to onset and progression of periodontitis is a complex relationship between oral bacteria and the host. The organisms most associated with severe periodontitis are the periodontal pathogens of the red complex: Tannerella forsythia, Treponema denticola and Porphyromonas gingivalis. These organisms express sialidases, which cleave sialic acid from host glycoproteins, and contribute to disease through various mechanisms. Here, we expressed and purified recombinant P. gingivalis sialidase SiaPG (PG_0352) and characterized its activity on a number of substrates, including host sialoglycoproteins and highlighting the inability to cleave diacetylated sialic acids - a phenomenon overcome by the NanS sialate-esterase from T. forsythia. Indeed SiaPG required NanS to maximize sialic acid harvesting from heavily O-acetylated substrates such as bovine salivary mucin, hinting at the possibility of interspecies cooperation in sialic acid release from host sources by these members of the oral microbiota. Activity of SiaPG and P. gingivalis was inhibited using the commercially available chemotherapeutic zanamivir, indicating its potential as a virulence inhibitor, which also inhibited sialic acid release from mucin, and was capable of inhibiting biofilm formation of P. gingivalis on oral glycoprotein sources. Zanamivir also inhibited attachment and invasion of oral epithelial cells by P. gingivalis and other periodontal pathogens, both in monospecies but also in multispecies infection experiments, indicating potential to suppress host-pathogen interactions of a mixed microbial community. This study broadens our understanding of the multifarious roles of bacterial sialidases in virulence, and indicates that their inhibition with chemotherapeutics could be a promising strategy for periodontitis therapy.