Rapid desensitization of the rat α7 nAChR is facilitated by the presence of a proline residue in the outer β-sheet

Thomas J. McCormack, Claudio Melis, José Colón, Elaine A. Gay, A. Mike, Robert Karoly, Patricia W. Lamb, Carla Molteni, Jerrel L. Yakel

Research output: Contribution to journalArticle

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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. The rat α7 nicotinic acetylcholine receptor (nAChR) desensitizes in the continual presence of high concentrations of agonist. Although the mechanism of desensitization is unknown, understanding its properties might be important in the design of therapeutics currently under development to treat a variety of neurodegenerative diseases and disorders; many of these compounds that target the α7 receptor are thought to potentiate responses by reducing desensitization. Here we have identified a critical proline residue near the middle of the β9 strand in the extracellular binding domain of the α7 nAChR which appears to markedly regulate desensitization. This part of the receptor is structurally important in that it physically links the critical C- and F-loops of the extracellular domain, within the outer β-sheet of the rat α7 nAChR, a region previously shown to affect receptor activation.

Original languageEnglish
Pages (from-to)4415-4429
Number of pages15
JournalJournal of Physiology
Volume588
Issue number22
DOIs
Publication statusPublished - Nov 2010

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Nicotinic Receptors
Proline
Serine
Threonine
Neurodegenerative Diseases
Psychologic Desensitization
Mutation
Molecular Dynamics Simulation
Hydroxyl Radical

ASJC Scopus subject areas

  • Physiology

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Rapid desensitization of the rat α7 nAChR is facilitated by the presence of a proline residue in the outer β-sheet. / McCormack, Thomas J.; Melis, Claudio; Colón, José; Gay, Elaine A.; Mike, A.; Karoly, Robert; Lamb, Patricia W.; Molteni, Carla; Yakel, Jerrel L.

In: Journal of Physiology, Vol. 588, No. 22, 11.2010, p. 4415-4429.

Research output: Contribution to journalArticle

McCormack, TJ, Melis, C, Colón, J, Gay, EA, Mike, A, Karoly, R, Lamb, PW, Molteni, C & Yakel, JL 2010, 'Rapid desensitization of the rat α7 nAChR is facilitated by the presence of a proline residue in the outer β-sheet', Journal of Physiology, vol. 588, no. 22, pp. 4415-4429. https://doi.org/10.1113/jphysiol.2010.195495
McCormack, Thomas J. ; Melis, Claudio ; Colón, José ; Gay, Elaine A. ; Mike, A. ; Karoly, Robert ; Lamb, Patricia W. ; Molteni, Carla ; Yakel, Jerrel L. / Rapid desensitization of the rat α7 nAChR is facilitated by the presence of a proline residue in the outer β-sheet. In: Journal of Physiology. 2010 ; Vol. 588, No. 22. pp. 4415-4429.
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AU - Melis, Claudio

AU - Colón, José

AU - Gay, Elaine A.

AU - Mike, A.

AU - Karoly, Robert

AU - Lamb, Patricia W.

AU - Molteni, Carla

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N2 - 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. The rat α7 nicotinic acetylcholine receptor (nAChR) desensitizes in the continual presence of high concentrations of agonist. Although the mechanism of desensitization is unknown, understanding its properties might be important in the design of therapeutics currently under development to treat a variety of neurodegenerative diseases and disorders; many of these compounds that target the α7 receptor are thought to potentiate responses by reducing desensitization. Here we have identified a critical proline residue near the middle of the β9 strand in the extracellular binding domain of the α7 nAChR which appears to markedly regulate desensitization. This part of the receptor is structurally important in that it physically links the critical C- and F-loops of the extracellular domain, within the outer β-sheet of the rat α7 nAChR, a region previously shown to affect receptor activation.

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