Research Departments & Organizations
Research in Carolyn Slayman’s laboratory uses the plasma-membrane H+-ATPase of yeast (Saccharomyces cerevisiae) as a simple model for studies on P-type pumps, a physiologically important family that includes the Na+,K+-, H+,K+-, and Ca2+-ATPases of animal cells. These pumps control the ionic composition of cells; many of them also serve as important drug targets (e.g., for cardiac glycosides in the case of Na+,K+-ATPase and anti-ulcer drugs in the case of gastric H+,K+-ATPase).
Specialized Terms: Genetics of Ion Transport
Extensive Research Description
Over the past several years, the Slayman laboratory has constructed a large collection of site-directed mutants and used them to explore structure-function relationships in key regions of the 100 kDa H+-ATPase. Results of particular significance include: (1) discovery of a 13-amino acid stretch in stalk segment 4 within which mutations disrupt the interaction between the catalytic domain of the ATPase (located in the cytoplasm) and the proton channel (embedded in the membrane); and (2) clear implication of one face of stalk segment 5 in the metabolic activation of the ATPase by glucose. Cysteine residues have now been introduced into both regions, and methods are being developed to track conformational changes by means of fluorescent sulfhydryl reagents. With a recently published high resolution structure of sarcoplasmic reticulum Ca2+-ATPase as a template, biophysical data from the H+-ATPase mutants should yield helpful insights into the molecular mechanism by which ATP hydrolysis is coupled to cation transport.
In parallel, the H+-ATPase is being used as a tool to probe the way in which newly synthesized plasma membrane proteins are delivered from the endoplasmic reticulum to the cell surface, as well as the quality control mechanisms that remove poorly folded or otherwise defective proteins. During the past year, a nested series of truncations has been constructed and has provided intriguing evidence for an oligomerization step at the C-terminus of the ATPase; oligomerization appears to occur early in the secretory pathway and to be essential for normal biogenesis.
YALE-UCL Collaborative London, United Kingdom (2011)
Dr. Slayman is on the Joint Strategy Committee for the Yale-UCL Collaborative, an alliance which will provide opportunities for high-level scientific research, clinical and educational collaboration across the institutions involved: Yale University, Yale School of Medicine, Yale-New Haven Hospital...
Tandem phosphorylation of Ser-911 and Thr-912 at the C terminus of yeast plasma membrane H+-ATPase leads to glucose-dependent activation.
Lecchi S, Nelson CJ, Allen KE, Swaney DL, Thompson KL, Coon JJ, Sussman MR, Slayman CW. Tandem phosphorylation of Ser-911 and Thr-912 at the C terminus of yeast plasma membrane H+-ATPase leads to glucose-dependent activation. The Journal Of Biological Chemistry 2007, 282:35471-81. 2007
Role of transmembrane segment M8 in the biogenesis and function of yeast plasma-membrane H(+)-ATPase.
Guerra G, Petrov VV, Allen KE, Miranda M, Pardo JP, Slayman CW. Role of transmembrane segment M8 in the biogenesis and function of yeast plasma-membrane H(+)-ATPase. Biochimica Et Biophysica Acta 2007, 1768:2383-92. 2007
Effects of C-terminal truncations on trafficking of the yeast plasma membrane H+-ATPase.
Mason AB, Allen KE, Slayman CW. Effects of C-terminal truncations on trafficking of the yeast plasma membrane H+-ATPase. The Journal Of Biological Chemistry 2006, 281:23887-98. 2006
Conformational changes of yeast plasma membrane H(+)-ATPase during activation by glucose: role of threonine-912 in the carboxy-terminal tail.
Lecchi S, Allen KE, Pardo JP, Mason AB, Slayman CW. Conformational changes of yeast plasma membrane H(+)-ATPase during activation by glucose: role of threonine-912 in the carboxy-terminal tail. Biochemistry 2005, 44:16624-32. 2005