Project: Research project

Project Details


DESCRIPTION (adapted from the applicant's abstract): The objective of this
proposal is to analyze the relationship between the structure and function
of Na,K-ATPase. The specific aims are: 1. Na,K-ATPase alpha/beta subunit
interaction. The applicant will attempt to determine which combinations of
alpha and beta subunit isoforms can associate by inserting epitope tags into
cDNAs encoding alpha and beta subunit isoforms and expressing the constructs
in mammalian cells. 2. Structure-function of the alpha subunit. The
applicant will develop a refined topographical map of the alpha subunit
polypeptide by creating a cysteine-less alpha subunit mutant. Cysteine
residues will be sequential introduced into predicted extracellular or
cytoplasmic loops of the cysteine-less mutant, and the topology of
individual cysteine tags determined using biotin-labeled thiol- or
sulfhydryl-reactive reagents. A second goal will be to identify sequences
within the alpha subunit that form the sodium binding site. 3. Genetic
analysis of Looptail mice:Function of the alpha2 subunit. The applicant
will attempt to determine whether Looptail (Lp) mice exhibit alterations in
the sequence or expression of the Na,K-ATPase alpha2 subunit (Atp1a2) gene.
4. Molecular analysis of the beta3 subunit. The applicant proposes to
further characterize the recently discovered Na,K-ATPase beta3 subunit. The
goal is to isolate and characterize cDNA and genomic DNA sequences for the
murine beta3 subunit and develop antibody probes for cell localization
Effective start/end date1/1/908/31/01


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