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":"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}
引用次数: 0
Abstract
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.