# CHARACTERIZATION OF TRANSVERSE TUBULE MG2+-ATPASE

> **NIH NIH R01** · UNIVERSITY OF CINCINNATI · 1990 · $89,617

## Abstract

The long term goals of the proposed research are to gain a better
understanding of the role of transverse tubules in the process of
excitation/contraction coupling in muscle and to understand how that
process is regulated by pharmacologically important and clinically
significant ions and drugs.  The transverse tubules play a pivotal role in
excitation/contraction coupling and abnormalities of the transverse tubules
may be important in muscular dystrophy.  These membranes have very
interesting pharmacological and biochemical characteristics, including a
preponderance of Ca2+ antagonist drug binding sites, a high level of
cholesterol, and a highly active, unique Mg2+-ATPase which may play a key
role in excitation/contraction coupling.  The specific aims are to
identify, purify, characterize in terms of possible physiological function,
and affinity label the Mg2+-ATPase.  Polyclonal antibodies will be raised
to the subunits of the Mg2+ -Atpase.  N-terminal sequencing of the purified
subunits as well as enzymatic and chemical fragmentation followed by
purified of peptides for sequencing will be performed.  From the protein
sequence obtained, oligonucleotide probes will be made to allow the
identification of the cDNA coding for the Mg2+ -ATPase and the entire amino
acid sequence deduced from the cDNA sequence.  A membrane spanning model
will be proposed based on hydropathy plots and labeling of sites on the
cytoplasmic face of the protein.  This ATPase will be compared structurally
and functionally to other ATPases.  The Mg2+ -ATPase may be a member of a
previously unknown class of ATPases, and structural comparisons will be
invaluable in the determination of essential and therefore conserved,
domains.  The techniques utilized in this work are centrifugation for
native gel electrophoresis and sucrose density centrifugation for isolation
of the enzyme, trypsin and cyanogen bromide fragmentation, reversed phase
HPLC, and amino acid analysis and gas phase sequencing for the
determination of the amino terminal sequence of the protein and generated
peptide fragments.  Techniques of cDNA cloning and sequencing will be used
to determine the entire sequence of the gene coding for the Mg2+ -ATPase.
This research will supply important information about a transverse tubule
component involved in excitation/contraction coupling which will lead to a
better understanding of this physiologically crucial process.

## Key facts

- **NIH application ID:** 3158635
- **Project number:** 5R01AR038576-02
- **Recipient organization:** UNIVERSITY OF CINCINNATI
- **SAM.gov UEI:** DZ4YCZ3QSPR5
- **Principal Investigator:** TERENCE LEE KIRLEY
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1990
- **Award amount:** $89,617
- **Award type:** 5
- **Project period:** 1989-03-01 → 1994-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3158635

## Citation

> US National Institutes of Health, RePORTER application 3158635, CHARACTERIZATION OF TRANSVERSE TUBULE MG2+-ATPASE (5R01AR038576-02). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3158635. Licensed CC0.

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