title: The osmotic activation of transporter ProP is tuned by both its C-terminal coiled-coil and osmotically induced changes in phospholipid composition.
abstract: Transporter ProP of Escherichia coli (ProPEc) senses extracellular osmolality and mediates osmoprotectant uptake when it is rising or high. A replica of the ProPEc C terminus (Asp468-Arg497) forms an intermolecular alpha-helical coiled-coil. This structure is implicated in the osmoregulation of intact ProPEc, in vivo. Like that from Corynebacterium glutamicum (ProPCg), the ProP orthologue from Agrobacterium tumefaciens (ProPAt) sensed and responded to extracellular osmolality after expression in E. coli. The osmotic activation profiles of all three orthologues depended on the osmolality of the bacterial growth medium, the osmolality required for activation rising as the growth osmolality approached 0.7 mol/kg. Thus, each could undergo osmotic adaptation. The proportion of cardiolipin in a polar lipid extract from E. coli increased with extracellular osmolality so that the osmolality activating ProPEc was a direct function of membrane cardiolipin content. Group A ProP orthologues (ProPEc, ProPAt) share the C-terminal coiled-coil domain and were activated at low osmolalities. Like variant ProPEc-R488I, in which the C-terminal coiled-coil is disrupted, ProPEc derivatives that lack the coiled-coil and Group B orthologue ProPCg required a higher osmolality to activate. The amplitude of ProPEc activation was reduced 10-fold in its deletion derivatives. The coiled-coil structure is not essential for osmotic activation of ProP per se. However, it tunes Group A orthologues to osmoregulate over a low osmolality range. Coiled-coil lesions may impair both coiled-coil formation and interaction of ProPEc with amplifier protein ProQ. Cardiolipin may contribute to ProP adaptation by altering bulk membrane properties or by acting as a ProP ligand.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
authors.forename: Yonit
authors.initials: Y
authors.lastname: Tsatskis
authors.forename: Jumana
authors.initials: J
authors.lastname: Khambati
authors.forename: Martina
authors.initials: M
authors.lastname: Dobson
authors.forename: Mikhail
authors.initials: M
authors.lastname: Bogdanov
authors.forename: William
authors.initials: W
authors.lastname: Dowhan
authors.forename: Janet M
authors.initials: JM
authors.lastname: Wood
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry; D001224:Aspartic Acid / Q000737:chemistry; D001665:Binding Sites; D001692:Biological Transport; D015153:Blotting, Western; D002308:Cardiolipins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D018388:Codon, Terminator; D003470:Culture Media / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism / Q000502:physiology*; D017353:Gene Deletion; D007477:Ions; D008024:Ligands; D008055:Lipids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D026901:Membrane Transport Proteins / Q000378:metabolism; D008956:Models, Chemical; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009994:Osmolar Concentration; D009995:Osmosis; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011188:Potassium / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051116:Receptors, Scavenger / Q000378:metabolism; D027981:Symporters / Q000378:metabolism / Q000502:physiology*
chemical_list: D002308:Cardiolipins; D018388:Codon, Terminator; D003470:Culture Media; D029968:Escherichia coli Proteins; D007477:Ions; D008024:Ligands; D008055:Lipids; D026901:Membrane Transport Proteins; D010743:Phospholipids; C079244:ProP protein, E coli; D051116:Receptors, Scavenger; D027981:Symporters; D001224:Aspartic Acid; D001120:Arginine; D011188:Potassium