Thanh-Nguyen Le, Thuan-Thien Dinh, Thuy-Dung Mai-Hoang, Ebrahim Razzazi-Fazeli, Hieu Tran-Van
{"title":"万年青中的丝氨酸蛋白酶抑制剂 3(Serpin3)选择性地与副溶血性弧菌 PirAvp 相互作用","authors":"Thanh-Nguyen Le, Thuan-Thien Dinh, Thuy-Dung Mai-Hoang, Ebrahim Razzazi-Fazeli, Hieu Tran-Van","doi":"10.1111/jfd.13935","DOIUrl":null,"url":null,"abstract":"Acute Hepatopancreatic Necrosis Disease (AHPND) represents a significant challenge in the field of shrimp aquaculture. This disease is primarily caused by <i>Vibrio parahaemolyticus</i> strains harbouring the pVA1 plasmid encoding the PirA<sup>vp</sup> and PirB<sup>vp</sup> toxins. To combat this epidemic and mitigate its devastating consequences, it is crucial to identify and characterize the receptors responsible for the binding of these pathogenic toxins. Our studied discovered that <i>Penaeus vannamei'</i>s Serine protease inhibitor 3 (<i>Pv</i>Serpin3) derived from shrimp hepatopancreatic tissues could bind to recombinant PirA<sup>vp</sup>, confirming its role as a novel PirA<sup>vp</sup>-binding protein (P<sub>A</sub>BP). Through comprehensive computational methods, we revealed two truncated PirA<sup>vp</sup>–binding proteins derived from <i>Pv</i>Serpin3 called Serpin3(13) and Serpin3(22), which had higher affinity to PirA<sup>vp</sup> than the full-length <i>Pv</i>Serpin3. The P<sub>A</sub>BP genes were amplified from a cDNA library that was reversed from total RNA extracted from shrimp, cloned and expressed in <i>Escherichia coli</i>. Three P<sub>A</sub>BP inclusion bodies were refolded to obtain the soluble form, and the recovery efficacy was found to be 100% for Serpin3 and Serpin3(13), while Serpin3(22) had a recovery efficacy of roundly 50%. Co-Immunoprecipitation (co-IP) and dot blot assays substantiated the interaction of these recombinant P<sub>A</sub>BPs with both recombinant PirA<sup>vp</sup> and VP<sub>AHPND</sub> (XN89)-producing natural toxins.","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.2000,"publicationDate":"2024-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Serine protease inhibitor 3 (Serpin3) from Penaeus vannamei selectively interacts with Vibrio parahaemolyticus PirAvp\",\"authors\":\"Thanh-Nguyen Le, Thuan-Thien Dinh, Thuy-Dung Mai-Hoang, Ebrahim Razzazi-Fazeli, Hieu Tran-Van\",\"doi\":\"10.1111/jfd.13935\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Acute Hepatopancreatic Necrosis Disease (AHPND) represents a significant challenge in the field of shrimp aquaculture. This disease is primarily caused by <i>Vibrio parahaemolyticus</i> strains harbouring the pVA1 plasmid encoding the PirA<sup>vp</sup> and PirB<sup>vp</sup> toxins. To combat this epidemic and mitigate its devastating consequences, it is crucial to identify and characterize the receptors responsible for the binding of these pathogenic toxins. Our studied discovered that <i>Penaeus vannamei'</i>s Serine protease inhibitor 3 (<i>Pv</i>Serpin3) derived from shrimp hepatopancreatic tissues could bind to recombinant PirA<sup>vp</sup>, confirming its role as a novel PirA<sup>vp</sup>-binding protein (P<sub>A</sub>BP). Through comprehensive computational methods, we revealed two truncated PirA<sup>vp</sup>–binding proteins derived from <i>Pv</i>Serpin3 called Serpin3(13) and Serpin3(22), which had higher affinity to PirA<sup>vp</sup> than the full-length <i>Pv</i>Serpin3. The P<sub>A</sub>BP genes were amplified from a cDNA library that was reversed from total RNA extracted from shrimp, cloned and expressed in <i>Escherichia coli</i>. Three P<sub>A</sub>BP inclusion bodies were refolded to obtain the soluble form, and the recovery efficacy was found to be 100% for Serpin3 and Serpin3(13), while Serpin3(22) had a recovery efficacy of roundly 50%. Co-Immunoprecipitation (co-IP) and dot blot assays substantiated the interaction of these recombinant P<sub>A</sub>BPs with both recombinant PirA<sup>vp</sup> and VP<sub>AHPND</sub> (XN89)-producing natural toxins.\",\"PeriodicalId\":15849,\"journal\":{\"name\":\"Journal of fish diseases\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2024-02-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of fish diseases\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1111/jfd.13935\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"FISHERIES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of fish diseases","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1111/jfd.13935","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"FISHERIES","Score":null,"Total":0}
Serine protease inhibitor 3 (Serpin3) from Penaeus vannamei selectively interacts with Vibrio parahaemolyticus PirAvp
Acute Hepatopancreatic Necrosis Disease (AHPND) represents a significant challenge in the field of shrimp aquaculture. This disease is primarily caused by Vibrio parahaemolyticus strains harbouring the pVA1 plasmid encoding the PirAvp and PirBvp toxins. To combat this epidemic and mitigate its devastating consequences, it is crucial to identify and characterize the receptors responsible for the binding of these pathogenic toxins. Our studied discovered that Penaeus vannamei's Serine protease inhibitor 3 (PvSerpin3) derived from shrimp hepatopancreatic tissues could bind to recombinant PirAvp, confirming its role as a novel PirAvp-binding protein (PABP). Through comprehensive computational methods, we revealed two truncated PirAvp–binding proteins derived from PvSerpin3 called Serpin3(13) and Serpin3(22), which had higher affinity to PirAvp than the full-length PvSerpin3. The PABP genes were amplified from a cDNA library that was reversed from total RNA extracted from shrimp, cloned and expressed in Escherichia coli. Three PABP inclusion bodies were refolded to obtain the soluble form, and the recovery efficacy was found to be 100% for Serpin3 and Serpin3(13), while Serpin3(22) had a recovery efficacy of roundly 50%. Co-Immunoprecipitation (co-IP) and dot blot assays substantiated the interaction of these recombinant PABPs with both recombinant PirAvp and VPAHPND (XN89)-producing natural toxins.
期刊介绍:
Journal of Fish Diseases enjoys an international reputation as the medium for the exchange of information on original research into all aspects of disease in both wild and cultured fish and shellfish. Areas of interest regularly covered by the journal include:
-host-pathogen relationships-
studies of fish pathogens-
pathophysiology-
diagnostic methods-
therapy-
epidemiology-
descriptions of new diseases