Alex S. Evers, M.D.
Henry Mallinckrodt Professor and Head
Computational and Molecular Biophysics Program
Anesthetics, photolabeling, mass spectrometry, proteomics
Molecular mechanisms of anesthetic action - Proteomics of GABAA receptors
My laboratory focuses on the protein chemistry of GABAA receptors to identify and characterize drug binding sites, sequence variants and post-translational modifications as well as to define the protein complexes in which GABAA receptors reside in vivo. A variety of biochemical and molecular biological approaches are applied to these problems, with particular emphasis on high-resolution mass spectrometry.
The lab has a longstanding interest in identifying the sites to which anesthetics bind to produce their effects. We have focused on anesthetic neurosteroids, using neurosteroid analogue photolabeling reagents to covalently label protein binding sites and identifying the sites of incorporation using protein mass spectrometry.
The lab has also pioneered methods for mass spectrometric methods to sequence the transmembrane segments of native GABAA receptor from picomole quantities of protein. Current directions in the lab include: 1) Studies of dynamic regulation of protein phosphorylation (particularly ion channels) in native brain using global (“shotgun”) mass spectrometric techniques and 2) Identification of GABAA-receptor associated proteins in brain using immunopurification coupled to mass spectrometry with label-free quantitation.
Chen ZW, Fuchs K, Sieghart W, Townsend RR, Evers AS. Deep amino acid sequencing of native brain GABAA receptors using high-resolution mass spectrometry. Mol Cell Proteomics 2011 (In Press). epub ahead of print PMID:21969609
Evers AS, Chen ZW, Manion BD, Han M, Jiang X, Darbandi-Tonkabon R, Kable T, Bracamontes J, Zorumski CF, Mennerick S, Steinbach JH, Covey DF. A Synthetic 18-Norsteroid distinguishes between two neuroactive steroid binding sites on GABAA receptors. J Pharmacol Exp Ther 2010 333(2): 404-13.
Darbandi-Tonkabon R, Manion B, Hastings WR, Craigen WJ, Akk G, Bracamontes JR, He Y, Sheiko TV, Steinbach JH, Mennerick SJ, Covey DF, Evers AS. Neuroactive steroid interactions with voltage-dependent anion channels: Relationship to GABAA receptor modulation and anesthesia. J Pharmacol Exp Ther 2004 308: 502-511.
Darbandi-Tonkabon R, Hastings WR, Zeng, CM, Akk G, Manion BD, Bracamontes JR, Steinbach JH, Mennerick SJ, Covey DF and Evers AS. Photoaffinity labeling with a neuroactive steroid analogue: 6-azi-pregnanolone labels voltage-dependent anion channel-1 in rat brain. J Biol Chem 2003 278: 13196-13206.
Evers AS, Berkowitz BA, d`Avignon DA. Correlation between the anesthetic effect of halothane and saturable binding in brain. Nature 1987 328: 157-160.
Last Updated: 1/19/2012 11:28:43 AM