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Inter- and Intra-Subunit Butanol/Isoflurane Sites of Action in the Human Glycine Receptor

Overview of attention for article published in Frontiers in Molecular Neuroscience, June 2016
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Title
Inter- and Intra-Subunit Butanol/Isoflurane Sites of Action in the Human Glycine Receptor
Published in
Frontiers in Molecular Neuroscience, June 2016
DOI 10.3389/fnmol.2016.00045
Pubmed ID
Authors

Mandy L. McCracken, Giorgio Gorini, Lindsay M. McCracken, R. Dayne Mayfield, R. Adron Harris, James R. Trudell

Abstract

Glycine receptors (GlyRs) mediate inhibitory neurotransmission and are targets for alcohols and anesthetics in brain. GlyR transmembrane (TM) domains contain critical residues for alcohol/anesthetic action: amino acid A288 in TM3 forms crosslinks with TM1 (I229) in the adjacent subunit as well as TM2 (S267) and TM4 (Y406, W407, I409, Y410) in the same subunit. We hypothesized that these residues may participate in intra-subunit and inter-subunit sites of alcohol/anesthetic action. The following double and triple mutants of GLRA1 cDNA (encoding human glycine receptor alpha 1 subunit) were injected into Xenopus laevis oocytes: I229C/A288C, I229C/A288C/C290S, A288C/Y406C, A288C/W407C, A288C/I409C, and A288C/Y410C along with the corresponding single mutants and wild-type GLRA1. Butanol (22 mM) or isoflurane (0.6 mM) potentiation of GlyR-mediated currents before and after application of the cysteine crosslinking agent HgCl2 (10 μM) was measured using two-electrode voltage clamp electrophysiology. Crosslinking nearly abolished butanol and isoflurane potentiation in the I229C/A288C and I229C/A288C/C290S mutants but had no effect in single mutants or wild-type. Crosslinking also inhibited butanol and isoflurane potentiation in the TM3-4 mutants (A288C/Y406C, A288C/W407C, A288C/I409C, A288C/Y410C) with no effect in single mutants or wild-type. We extracted proteins from oocytes expressing I229C/288C, A288C/Y410C, or wild-type GlyRs, used mass spectrometry to verify their expression and possible inter-subunit dimerization, plus immunoblotting to investigate the biochemical features of proposed crosslinks. Wild-type GlyR subunits measured about 50 kDa; after crosslinking, the dimeric/monomeric 100:50 kDa band ratio was significantly increased in I229C/288C but not A288C/Y410C mutants or wild-type, providing support for TM1-3 inter-subunit and TM3-4 intra-subunit crosslinking. A GlyR homology model based on the GluCl template provides further evidence for a multi-site model for alcohol/anesthetic interaction with human GLRA1.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 11 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 11 100%

Demographic breakdown

Readers by professional status Count As %
Student > Master 2 18%
Student > Ph. D. Student 2 18%
Unspecified 1 9%
Student > Doctoral Student 1 9%
Student > Bachelor 1 9%
Other 2 18%
Unknown 2 18%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 3 27%
Veterinary Science and Veterinary Medicine 1 9%
Unspecified 1 9%
Pharmacology, Toxicology and Pharmaceutical Science 1 9%
Agricultural and Biological Sciences 1 9%
Other 2 18%
Unknown 2 18%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 06 July 2016.
All research outputs
#14,855,186
of 22,877,793 outputs
Outputs from Frontiers in Molecular Neuroscience
#1,671
of 2,890 outputs
Outputs of similar age
#212,288
of 352,714 outputs
Outputs of similar age from Frontiers in Molecular Neuroscience
#15
of 29 outputs
Altmetric has tracked 22,877,793 research outputs across all sources so far. This one is in the 33rd percentile – i.e., 33% of other outputs scored the same or lower than it.
So far Altmetric has tracked 2,890 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.7. This one is in the 36th percentile – i.e., 36% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 352,714 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 37th percentile – i.e., 37% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 29 others from the same source and published within six weeks on either side of this one. This one is in the 44th percentile – i.e., 44% of its contemporaries scored the same or lower than it.