title: Lipids and topological rules of membrane protein assembly: balance between long and short range lipid-protein interactions.
abstract: The N-terminal six-transmembrane domain (TM) bundle of lactose permease of Escherichia coli is uniformly inverted when assembled in membranes lacking phosphatidylethanolamine (PE). Inversion is dependent on the net charge of cytoplasmically exposed protein domains containing positive and negative residues, net charge of the membrane surface, and low hydrophobicity of TM VII acting as a molecular hinge between the two halves of lactose permease (Bogdanov, M., Xie, J., Heacock, P., and Dowhan, W. (2008) J. Cell Biol. 182, 925-935). Net neutral lipids suppress the membrane translocation potential of negatively charged amino acids, thus increasing the cytoplasmic retention potential of positively charged amino acids. Herein, TM organization of sucrose permease (CscB) and phenylalanine permease (PheP) as a function of membrane lipid composition was investigated to extend these principles to other proteins. For CscB, topological dependence on PE only becomes evident after a significant increase in the net negative charge of the cytoplasmic surface of the N-terminal TM bundle. High negative charge is required to overcome the thermodynamic block to inversion due to the high hydrophobicity of TM VII. Increasing the positive charge of the cytoplasmic surface of the N-terminal TM hairpin of PheP, which is misoriented in PE-lacking cells, favors native orientation in the absence of PE. PheP and CscB also display co-existing dual topologies dependent on changes in the charge balance between protein domains and the membrane lipids. Therefore, the topology of both permeases is dependent on PE. However, CscB topology is governed by thermodynamic balance between opposing lipid-dependent electrostatic and hydrophobic interactions.
authors: Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston, Texas 77030, USA.
authors: Heidi
authors: H
authors: Vitrac
authors: Mikhail
authors: M
authors: Bogdanov
authors: Phil
authors: P
authors: Heacock
authors: William
authors: W
authors: Dowhan
mesh_terms: D026921:Amino Acid Transport Systems, Neutral / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000378:metabolism
chemical_list: D026921:Amino Acid Transport Systems, Neutral; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008563:Membrane Lipids; D026901:Membrane Transport Proteins; D010714:Phosphatidylethanolamines; C069320:phenylalanine-specific permease, E coli; C056379:cscB protein, E coli; C021499:lactose permease