Dear Editor,The orientation of a membrane protein can provide important information about its interaction partners,function,and mechanism of action.In addition to traditional biochemical assays,new methods have been d...Dear Editor,The orientation of a membrane protein can provide important information about its interaction partners,function,and mechanism of action.In addition to traditional biochemical assays,new methods have been developed to reveal the orientation of membrane proteins.For example,computer programs have been developed to predict membrane protein topology but often have inaccuracies(Ott and Lingappa,2002;Schwacke et al.,2003).A widely used method is the fluorescence protease protection(FPP)assay(Lomize et al.,2006;Lorenz et al.,2008;White et al.,2015).However,it often fails to give good results for assays of plant membrane proteins.Here,we developed a simpler method based on release of a fluorescent protein tag(RFT)by the tobacco etch virus(TEV)protease to overcome the shortcomings of FPP and analyze the orientation of membrane proteins in plants.展开更多
基金supported by grants from the Fundamental Research Funds for the Central Universities(2022BLRD14)the National Natural Science Foundation of China(nos.32070696 and 31570182).
文摘Dear Editor,The orientation of a membrane protein can provide important information about its interaction partners,function,and mechanism of action.In addition to traditional biochemical assays,new methods have been developed to reveal the orientation of membrane proteins.For example,computer programs have been developed to predict membrane protein topology but often have inaccuracies(Ott and Lingappa,2002;Schwacke et al.,2003).A widely used method is the fluorescence protease protection(FPP)assay(Lomize et al.,2006;Lorenz et al.,2008;White et al.,2015).However,it often fails to give good results for assays of plant membrane proteins.Here,we developed a simpler method based on release of a fluorescent protein tag(RFT)by the tobacco etch virus(TEV)protease to overcome the shortcomings of FPP and analyze the orientation of membrane proteins in plants.
