title: Slow alpha helix formation during folding of a membrane protein.
abstract: Very little is known about the folding of proteins within biological membranes. A "two-stage" model has been proposed on thermodynamic grounds for the folding of alpha helical, integral membrane proteins, the first stage of which involves formation of transmembrane alpha helices that are proposed to behave as autonomous folding domains. Here, we investigate alpha helix formation in bacteriorhodopsin and present a time-resolved circular dichroism study of the slow in vitro folding of this protein. We show that, although some of the protein's alpha helices form early, a significant part of the protein's secondary structure appears to form late in the folding process. Over 30 amino acids, equivalent to at least one of bacteriorhodopsin's seven transmembrane segments, slowly fold from disordered structures to alpha helices with an apparent rate constant of about 0.012 s-1 at pH 6 or 0.0077 s-1 at pH 8. This is a rate-limiting step in protein folding, which is dependent on the pH and the composition of the lipid bilayer.
authors.affiliation: Department of Biochemistry, Imperial College of Science, Technology, and Medicine, London, U.K.
authors.forename: M L
authors.initials: ML
authors.lastname: Riley
authors.forename: B A
authors.initials: BA
authors.lastname: Wallace
authors.forename: S L
authors.initials: SL
authors.lastname: Flitsch
authors.forename: P J
authors.initials: PJ
authors.lastname: Booth
mesh_terms: D001436:Bacteriorhodopsins / Q000378:metabolism*; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D005470:Fluorometry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry*; D008956:Models, Chemical; D010742:Phospholipid Ethers / Q000378:metabolism; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013816:Thermodynamics
chemical_list: C052298:1,2-dihexadecyl-sn-glycero-3-phosphocholine; D008081:Liposomes; D008565:Membrane Proteins; D010742:Phospholipid Ethers; D001436:Bacteriorhodopsins; D004134:Dimyristoylphosphatidylcholine