Reconstitution of outer membrane protein assembly from purified components.
Beta-barrel membrane proteins in Gram-negative bacteria, mitochondria, and chloroplasts are assembled by highly conserved multi-protein complexes. The mechanism by which these molecular machines fold and insert their substrates is poorly understood. It has not been possible to dissect the folding and insertion pathway because the process has not been reproduced in a biochemical system. We purified the components that fold and insert Escherichia coli outer membrane proteins and reconstituted beta-barrel protein assembly in proteoliposomes using the enzymatic activity of a protein substrate to report on its folding state. The assembly of this protein occurred without an energy source but required a soluble chaperone in addition to the multi-protein assembly complex.
Hagan
Christine L
CL
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
Kim
Seokhee
S
Kahne
Daniel
D
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D008074:Lipoproteins / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry*; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D019696:Peptidylprolyl Isomerase / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011510:Proteolipids / Q000737:chemistry
D001425:Bacterial Outer Membrane Proteins; C503914:BamA protein, E coli; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D008074:Lipoproteins; D008081:Liposomes; D018832:Molecular Chaperones; D046912:Multiprotein Complexes; D011510:Proteolipids; C499588:ompT protein, E coli; C020485:proteoliposomes; D010447:Peptide Hydrolases; C490900:SurA protein, E coli; D019696:Peptidylprolyl Isomerase