title: Assembly of eukaryotic class III (N-out, C-in) membrane proteins into the Escherichia coli cytoplasmic membrane.
abstract: Class III membrane proteins lack cleavable signal peptides but adopt an N-out, C-in topology with respect to their native membranes. We have analysed the fate of two eukaryotic class III plasma membrane proteins, human erythrocyte glycophorin C and influenza A virus M2 protein, in Escherichia coli. The N-terminal domains of both proteins were efficiently localised to the extracytoplasmic side of the bacterial cytoplasmic membrane. When beta-lactamase was fused to the C-terminus of glycophorin C it was localised to the cytoplasm, and protease treatment of spheroplasts caused a reduction in size of the fusion protein consistent with glycophorin C adopting its native topology in E. coli.
authors.affiliation: Microbial Genetics Group, School of Biological Sciences, University of Sussex, Falmer, Brighton, UK.
authors.forename: E S
authors.initials: ES
authors.lastname: Hennessey
authors.forename: L
authors.initials: L
authors.lastname: Hashemzadeh-Bonehi
authors.forename: L A
authors.initials: LA
authors.lastname: Hunt
authors.forename: J K
authors.initials: JK
authors.lastname: Broome-Smith
mesh_terms: D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006021:Glycophorins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009980:Influenza A virus / Q000235:genetics*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000086082:Viroporin Proteins
chemical_list: D018076:DNA, Complementary; D006021:Glycophorins; D008565:Membrane Proteins; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins