The long-term goal of this project is an understanding of structure-function relationships in the Na,K-pump. The investigator has proposed studies in protein chemistry and affinity labeling that are a straightforward extension of his previous work. The cation complexation site within the catalytic alpha subunit of the pump will be identified by affinity labeling with positively charged carboxyl-reactive compounds (Specific Aim 1). The isothiocyanate-reactive lysine in the major cytoplasmic domain (Lys-501) will be used as an anchor in cross-linking experiments designed to identify those residues involved with nucleotide binding and energy transduction (Aim 2). Additional cross-linking and photolabeling will be used in right-side-out microsomes to test directly the proposed membrane topology derived from hydropathy analysis (Aim 3). Finally, photoactive derivatives of the cardiotonic steroids will be used to identify amino acids that contribute to glycoside binding (Aim 4).
| Clifford, Rebecca J; Kaplan, Jack H (2013) Human breast tumor cells are more resistant to cardiac glycoside toxicity than non-tumorigenic breast cells. PLoS One 8:e84306 |
| Tokhtaeva, Elmira; Clifford, Rebecca J; Kaplan, Jack H et al. (2012) Subunit isoform selectivity in assembly of Na,K-ATPase ?-? heterodimers. J Biol Chem 287:26115-25 |
| Clifford, Rebecca J; Kaplan, Jack H (2009) Regulation of Na,K-ATPase subunit abundance by translational repression. J Biol Chem 284:22905-15 |
| Clifford, Rebecca J; Kaplan, Jack H (2008) beta-Subunit overexpression alters the stoicheometry of assembled Na-K-ATPase subunits in MDCK cells. Am J Physiol Renal Physiol 295:F1314-23 |
| Bystriansky, Jason S; Kaplan, Jack H (2007) Sodium pump localization in epithelia. J Bioenerg Biomembr 39:373-8 |
| Laughery, Melissa D; Clifford, Rebecca J; Chi, Yiqing et al. (2007) Selective basolateral localization of overexpressed Na-K-ATPase beta1- and beta2- subunits is disrupted by butryate treatment of MDCK cells. Am J Physiol Renal Physiol 292:F1718-25 |
| Laughery, Melissa; Todd, Matthew; Kaplan, Jack H (2004) Oligomerization of the Na,K-ATPase in cell membranes. J Biol Chem 279:36339-48 |
| Geibel, Sven; Kaplan, Jack H; Bamberg, Ernst et al. (2003) Conformational dynamics of the Na+/K+-ATPase probed by voltage clamp fluorometry. Proc Natl Acad Sci U S A 100:964-9 |
| Costa, Charles J; Gatto, Craig; Kaplan, Jack H (2003) Interactions between Na,K-ATPase alpha-subunit ATP-binding domains. J Biol Chem 278:9176-84 |
| Kaplan, Jack H (2002) Biochemistry of Na,K-ATPase. Annu Rev Biochem 71:511-35 |
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