title: Rapid desensitization of the rat α7 nAChR is facilitated by the presence of a proline residue in the outer β-sheet.
abstract: The rat α7 nicotinic acetylcholine receptor (nAChR) has a proline residue near the middle of the β9 strand. The replacement of this proline residue at position 180 (P180) by either threonine (α7-P180T) or serine (α7-P180S) slowed the onset of desensitization dramatically, with half-times of ~930 and 700 ms, respectively, compared to 90 ms for the wild-type receptor. To investigate the importance of the hydroxyl group on the position 180 side-chains, the mutant receptors α7-P180Y and α7-P180F were studied and showed half-times of desensitization of 650 and 160 ms, respectively. While a position 180 side-chain OH group may contribute to the slow desensitization rates, α7-P180S and α7-P180V resulted in receptors with similar desensitization rates, suggesting that increased backbone to backbone H bonding expected in the absence of proline at position 180 would likely exert a great effect on desensitization. Single channel recordings indicated that for the α7-P180T receptor there was a significantly reduced closed time without any change in single channel conductance (as compared to wild-type). Kinetic simulations indicated that all changes observed for the mutant channel behaviour were reproduced by decreasing the rate of desensitization, and increasing the microscopic affinity to resting receptors. Molecular dynamics (MD) simulations on a homology model were used to provide insight into likely H bond interactions within the outer β-sheet that occur when the P180 residue is mutated. All mutations analysed increased about twofold the predicted number of H bonds between the residue at position 180 and the backbone of the β10 strand. Moreover, the α7-P180T and α7-P180S mutations also formed some intrastrand H bonds along the β9 strand, although H bonding of the OH groups of the threonine or serine side-chains was predicted to be infrequent. Our results indicate that rapid desensitization of the wild-type rat α7 nAChR is facilitated by the presence of the proline residue within the β9 strand.
authors.affiliation: Laboratory of Neurobiology, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, NC 27709, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: McCormack
authors.forename: Claudio
authors.initials: C
authors.lastname: Melis
authors.forename: José
authors.initials: J
authors.lastname: Colón
authors.forename: Elaine A
authors.initials: EA
authors.lastname: Gay
authors.forename: Arpad
authors.initials: A
authors.lastname: Mike
authors.forename: Robert
authors.initials: R
authors.lastname: Karoly
authors.forename: Patricia W
authors.initials: PW
authors.lastname: Lamb
authors.forename: Carla
authors.initials: C
authors.lastname: Molteni
authors.forename: Jerrel L
authors.initials: JL
authors.lastname: Yakel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002642:Chick Embryo; D006224:Cricetinae; D003412:Cricetulus; D018360:Crystallography, X-Ray; D005260:Female; D015640:Ion Channel Gating / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D011392:Proline / Q000737:chemistry* / Q000235:genetics*; D017433:Protein Structure, Secondary; D051381:Rats; D011978:Receptors, Nicotinic / Q000737:chemistry* / Q000235:genetics*; D013997:Time Factors; D014982:Xenopus laevis; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: C579167:Chrna7 protein, human; C579165:Chrna7 protein, rat; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D011392:Proline