Ludo L J Schoenmakers, Max J den Uijl, Jelle Postma, Tim A P van den Akker, Wilhelm T S Huck, Arnold J. M. Driessen
{"title":"合成细胞模型中 SecYEG 介导的转运","authors":"Ludo L J Schoenmakers, Max J den Uijl, Jelle Postma, Tim A P van den Akker, Wilhelm T S Huck, Arnold J. M. Driessen","doi":"10.1093/synbio/ysae007","DOIUrl":null,"url":null,"abstract":"\n Giant unilamellar vesicles (GUVs) provide a powerful model compartment for synthetic cells. However, a key challenge is the incorporation of membrane proteins that allow for transport, energy transduction, compartment growth and division. Here, we have successfully incorporated the membrane protein complex SecYEG – the key bacterial translocase that is essential for the incorporation of newly synthesized membrane proteins – in GUVs. Our method consists of fusion of small unilamellar vesicles (SUVs) containing reconstituted SecYEG into GUVs, thereby forming SecGUVs. These are suitable for large scale experiments while maintaining a high protein:lipid ratio. We demonstrate that incorporation of SecYEG into GUVs does not inhibit its translocation efficiency. Robust membrane protein functionalized proteo-GUVs are promising and flexible compartments for use in the formation and growth of synthetic cells.","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2000,"publicationDate":"2024-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"SecYEG-mediated Translocation in a Model Synthetic Cell\",\"authors\":\"Ludo L J Schoenmakers, Max J den Uijl, Jelle Postma, Tim A P van den Akker, Wilhelm T S Huck, Arnold J. M. Driessen\",\"doi\":\"10.1093/synbio/ysae007\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"\\n Giant unilamellar vesicles (GUVs) provide a powerful model compartment for synthetic cells. However, a key challenge is the incorporation of membrane proteins that allow for transport, energy transduction, compartment growth and division. Here, we have successfully incorporated the membrane protein complex SecYEG – the key bacterial translocase that is essential for the incorporation of newly synthesized membrane proteins – in GUVs. Our method consists of fusion of small unilamellar vesicles (SUVs) containing reconstituted SecYEG into GUVs, thereby forming SecGUVs. These are suitable for large scale experiments while maintaining a high protein:lipid ratio. We demonstrate that incorporation of SecYEG into GUVs does not inhibit its translocation efficiency. Robust membrane protein functionalized proteo-GUVs are promising and flexible compartments for use in the formation and growth of synthetic cells.\",\"PeriodicalId\":22158,\"journal\":{\"name\":\"Synthetic Biology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.2000,\"publicationDate\":\"2024-05-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Synthetic Biology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1093/synbio/ysae007\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"Agricultural and Biological Sciences\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Synthetic Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/synbio/ysae007","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Agricultural and Biological Sciences","Score":null,"Total":0}
SecYEG-mediated Translocation in a Model Synthetic Cell
Giant unilamellar vesicles (GUVs) provide a powerful model compartment for synthetic cells. However, a key challenge is the incorporation of membrane proteins that allow for transport, energy transduction, compartment growth and division. Here, we have successfully incorporated the membrane protein complex SecYEG – the key bacterial translocase that is essential for the incorporation of newly synthesized membrane proteins – in GUVs. Our method consists of fusion of small unilamellar vesicles (SUVs) containing reconstituted SecYEG into GUVs, thereby forming SecGUVs. These are suitable for large scale experiments while maintaining a high protein:lipid ratio. We demonstrate that incorporation of SecYEG into GUVs does not inhibit its translocation efficiency. Robust membrane protein functionalized proteo-GUVs are promising and flexible compartments for use in the formation and growth of synthetic cells.