Site-directed mutagenesis of yeast V-ATPase subunit d

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dc.contributor.advisor Parra-Belky, Karlett J. en_US Owegi, Margaret en_US 2011-06-03T19:40:30Z 2011-06-03T19:40:30Z 2005 en_US 2005
dc.identifier LD2489.Z78 2005 .O94 en_US
dc.description.abstract V-ATPases are enzymes found in all eukaryotic cells. They are organized into a peripheral membrane complex (V1) and an integral membrane complex (V0). VI is responsible for ATP hydrolysis and generates the energy used by Vo to pump protons from the cytosol into the vacuole. Subunit d is a component of Vo possibly located at the interface between V 1 and V. in the V-ATPase complex. We hypothesize that subunit d could be involved in the structural and functional coupling of VI and Vo. This was tested by generating point mutations along the open reading frame of subunit d from yeast. The mutations F94A, H128A, D173A, D217A, D261A, E317A, W325A, E328A and C329A, all in conserved regions of the protein sequence, were characterized by examining their growth phenotype and by assessing their ATPase specific activity, proton transport and V1Vo assembly in purified vacuolar membranes. The mutations E317A, W325A, E328A and C329A had reduced ATPase and proton transport activities. In addition, V1Vo assembly was compromised by the mutation W325A. Our results suggest that residues at the carboxyl-end of subunit d are important for ATPase activity, proton pumping and V1Vo assembly at the membrane.
dc.description.sponsorship Department of Chemistry
dc.format.extent vi, 69 leaves : ill. (some col.) ; 28 cm. en_US
dc.source Virtual Press en_US
dc.subject.lcsh Adenosine triphosphatase -- Structure. en_US
dc.subject.lcsh Site-specific mutagenesis. en_US
dc.title Site-directed mutagenesis of yeast V-ATPase subunit d en_US
dc.title.alternative Site directed mutagenesis of yeast vacuolar adenosine triphosphatase subunit d en_US Thesis (M.S.)
dc.identifier.cardcat-url en_US

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  • Master's Theses [5454]
    Master's theses submitted to the Graduate School by Ball State University master's degree candidates in partial fulfillment of degree requirements.

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