Expression of membrane proteins at the Escherichia coli membrane for structural studies.
Structural biology of membrane proteins is often limited by the first steps in obtaining sufficient yields of proteins because native sources are seldom. Heterologous systems like bacteria are then commonly employed for membrane protein over-expression. Escherichia coli is the main bacterial host used. However, overproduction of a foreign membrane protein at a non-physiological level is usually toxic for cells or leads to inclusion body formation. Those effects can be reduced by optimizing the cell growth conditions, choosing the suitable bacterial strain and expression vector, and finally co-expressing the target protein and the b-subunit of E. coli adenosine triphosphate (ATP)-synthase, which triggers the proliferation of intracytoplasmic membranes. This chapter is devoted to help the experimenter in choosing the appropriate plasmid/bacterial host combination for optimizing the amount of the target membrane protein produced in its correct folded state.
Zoonens
Manuela
M
Université Paris, Institut de Biologie Physico-Chimique, France.
Miroux
Bruno
B
D025181:ATP Synthetase Complexes / Q000235:genetics; D000818:Animals; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D009154:Mutation; D011401:Promoter Regions, Genetic
D008565:Membrane Proteins; D025181:ATP Synthetase Complexes