Research in the Booth Group
Research in the Booth lab is directed at understanding structure-function relationships in proteins. Our primary model system for these studies utilizes the expression of a membrane protein, the epithelial sodium channel (ENaC), in a yeast host system. In this system, ENaC expression induces a salt-sensitive phenotype of growth inhibition. In humans, ENaC is rate-limiting for sodium reabsorption in the kidneys and essential for maintaining electrolyte balance. Gain- and loss-in-function genetic mutations within ENaC lead to hyper- and hypotension, respectively.
Research students in the Booth lab will gain experience in a diverse set of biochemistry laboratory procedures and techniques including:
- Cell Culture
- Polymerase Chain Reactions (PCR)
- Protein and Nucleic Acid Purification
- Gene Cloning
- Gel Electrophoresis/Western Blotting
Initially, traditional molecular biology techniques (i.e. PCR, plasmid isolation, etc.) will be used to generate random mutations in targeted regions of ENaC, which will be screened for salt sensitivity in yeast. Yeast cells expressing mutant ENaCs that result in a reverse of the salt sensitive phenotype will be subjected to further analysis including isolation of the mutant ENaC, bioinformatic analysis of the mutant’s sequence to identify the critical residue(s)/region, and advanced characterization of the mutant ENaC’s properties.
Downs, K.P. ‡, YingJia, S., Pasquali, A., Beldorth, I., Savage, M., Gallier, K. ‡, Garcia, T., Booth, R.E.*, and Walter, RB. Characterization of telomeres and telomerase expression in Xiphophorus. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 2012, 155(1), 89-94.
Perez, A.N., Zxhang, Z., Oehlers, L., Booth, R.E., Walter, R.B., and David, W.M.* Proteomic Analyses of the Gordon-Kosswig Melanoma Model. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 2012, 155(1), 81-8.
Zhang, Z., Wells, M.C., Boswell, M.G.†, Beldorth, I., Kirk, L.M.‡, Wang, Y., Wang, S., Savage, M., Walter, R.B., and Booth, R.E.* Identification of Robust Hypoxia Biomarker Candidates from Fin of Medaka (Oryzias latipes). Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 2012, 155(1), 11-7.
Boswell, M.G. †, Kirk, L. †, Wells, M.C., Ju, Z., Zhang,Z., Booth,R.E., and Walter, R.W.* Comparison of Gene Expression Responses to Hypoxia in Viviparous (Xiphophorus) and Oviparous (Oryzias) Fishes Using a Medaka Microarray. Comp. Biochem. Phys. Part C: Tox. & Pharm. 2009,149(2), 258-265.
Staruschenko A., Pochynyuk O., Booth, R.E., Stockand J.D.* Insight Towards Epithelial Na+ Channel (ENaC) Mechanism Revealed by the Acid-sensing Ion Channel (ASIC) Structure. IUBMB Life 2008, 60(9), 620-8.
McVey, W.J., Matthews, B., Motley, D.M., Linse, K.D., Blass, D.P. †, Booth, R.E., and Feakes, D.A.* Investigation of the Interactions of Polyhedral Borane Anions with Serum Albumins. J. Inorg. Biochem. 2008,102(4), 943-51.
Staruschenko, A.*, Adams, E., Booth, R.E., Stockand, J.D. Epithelial Na+ Channel Subunit Stoichiometry. Biophys Journal 2005, 88(6), 3966-75.
Staruschenko, A., Medina, J.L., Patel, P., Shapiro, M.S., Booth, R.E.*, and Stockand, J.D. Fluorescence Resonance Energy Transfer Analysis of Subunit Stoichiometry of the Epithelial Na+ Channel. Journal of Biological Chemistry 2004, 25:279(26), 27729-34.
Tong, Q., Booth, R.E.*, Worrell, R.T. and Stockand, J.D. Regulation of Na+ Transport by Aldosterone: Signaling Convergence and Cross-talk Between the PI3-K and MAPK 1/2 Cascades. American Journal Physiology 2004, 286(6), F1232-8.
Booth, R.E.,Lovell, S.C., Misquitta, S.A., and Bateman, Jr R.C.* Human Glutaminyl Cyclase and Bacterial Zinc Aminopeptidase Share a Common Fold and Active Site. BMC Biology 2004, 2(1), 2