Marta Sanz-Gaitero, Vincent De Maesschalck, Ankur Patel, Hannelore Longin, Vera Van Noort, Lorena Rodriguez-Rubio, Michael van Ryne, Katarzyna Danis-Wlodarczyk, Zuzanna Drulis-Kawa, Stephane Mesnage, Mark van Raaij, Rob Lavigne
{"title":"一种新的噬菌体编码喃喃苷酶 KTN6 Gp46 的结构和生化特征。","authors":"Marta Sanz-Gaitero, Vincent De Maesschalck, Ankur Patel, Hannelore Longin, Vera Van Noort, Lorena Rodriguez-Rubio, Michael van Ryne, Katarzyna Danis-Wlodarczyk, Zuzanna Drulis-Kawa, Stephane Mesnage, Mark van Raaij, Rob Lavigne","doi":"10.1089/phage.2023.0040","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Endolysins are phage-encoded lytic enzymes that degrade bacterial peptidoglycan at the end of phage lytic cycles to release new phage particles. These enzymes are being explored as an alternative to small-molecule antibiotics.</p><p><strong>Methods: </strong>The crystal structure of KTN6 Gp46 was determined and compared with a ColabFold model. Cleavage specificity was examined using a peptidoglycan digest and reversed-phase high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS).</p><p><strong>Results: </strong>The structure of KTN6 Gp46 could be determined at 1.4 Å resolution, and key differences in loops of the putative peptidoglycan binding domain were identified in comparison with its closest known homologue, the endolysin of phage SPN1S. Reversed-phase HPLC/MS analysis of the reaction products following peptidoglycan digestion confirmed the muramidase activity of Gp46, consistent with structural predictions.</p><p><strong>Conclusion: </strong>These insights into the structure and function of endolysins further expand the toolbox for endolysin engineering and explore their potential in enzyme-based antibacterial design strategies.</p>","PeriodicalId":74428,"journal":{"name":"PHAGE (New Rochelle, N.Y.)","volume":"5 2","pages":"53-62"},"PeriodicalIF":0.0000,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11304755/pdf/","citationCount":"0","resultStr":"{\"title\":\"Structural and Biochemical Characterization of a New Phage-Encoded Muramidase, KTN6 Gp46.\",\"authors\":\"Marta Sanz-Gaitero, Vincent De Maesschalck, Ankur Patel, Hannelore Longin, Vera Van Noort, Lorena Rodriguez-Rubio, Michael van Ryne, Katarzyna Danis-Wlodarczyk, Zuzanna Drulis-Kawa, Stephane Mesnage, Mark van Raaij, Rob Lavigne\",\"doi\":\"10.1089/phage.2023.0040\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Endolysins are phage-encoded lytic enzymes that degrade bacterial peptidoglycan at the end of phage lytic cycles to release new phage particles. These enzymes are being explored as an alternative to small-molecule antibiotics.</p><p><strong>Methods: </strong>The crystal structure of KTN6 Gp46 was determined and compared with a ColabFold model. Cleavage specificity was examined using a peptidoglycan digest and reversed-phase high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS).</p><p><strong>Results: </strong>The structure of KTN6 Gp46 could be determined at 1.4 Å resolution, and key differences in loops of the putative peptidoglycan binding domain were identified in comparison with its closest known homologue, the endolysin of phage SPN1S. Reversed-phase HPLC/MS analysis of the reaction products following peptidoglycan digestion confirmed the muramidase activity of Gp46, consistent with structural predictions.</p><p><strong>Conclusion: </strong>These insights into the structure and function of endolysins further expand the toolbox for endolysin engineering and explore their potential in enzyme-based antibacterial design strategies.</p>\",\"PeriodicalId\":74428,\"journal\":{\"name\":\"PHAGE (New Rochelle, N.Y.)\",\"volume\":\"5 2\",\"pages\":\"53-62\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-06-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11304755/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"PHAGE (New Rochelle, N.Y.)\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1089/phage.2023.0040\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/6/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"PHAGE (New Rochelle, N.Y.)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/phage.2023.0040","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/6/1 0:00:00","PubModel":"eCollection","JCR":"","JCRName":"","Score":null,"Total":0}
Structural and Biochemical Characterization of a New Phage-Encoded Muramidase, KTN6 Gp46.
Background: Endolysins are phage-encoded lytic enzymes that degrade bacterial peptidoglycan at the end of phage lytic cycles to release new phage particles. These enzymes are being explored as an alternative to small-molecule antibiotics.
Methods: The crystal structure of KTN6 Gp46 was determined and compared with a ColabFold model. Cleavage specificity was examined using a peptidoglycan digest and reversed-phase high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS).
Results: The structure of KTN6 Gp46 could be determined at 1.4 Å resolution, and key differences in loops of the putative peptidoglycan binding domain were identified in comparison with its closest known homologue, the endolysin of phage SPN1S. Reversed-phase HPLC/MS analysis of the reaction products following peptidoglycan digestion confirmed the muramidase activity of Gp46, consistent with structural predictions.
Conclusion: These insights into the structure and function of endolysins further expand the toolbox for endolysin engineering and explore their potential in enzyme-based antibacterial design strategies.
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