STUDIES OF PURIFIED SODIUM CHANNELS FROM SARCOLEMMA

Project Details

Description

This proposal describes a continuation of our present work on the molecular
characterization of the voltage-sensitive sodium channel from mammalian
muscle. As in the past grant period, we will use a multidisciplinary
approach incorporating biochemical, biophysical and immunological
techniques to further probe the structure and function of this unique
molecule.

The sodium channel will be purified from rat and rabbit skeletal muscle.
The relationship between the large glycoprotein and the small subunit(s)
will be probed with monoclonal antibodies, peptide mapping and
immunoprecipitation. The large subunit will be studied in detail, with
analysis of amino acid composition, N-terminal sequence, and sugar content
and composition. Peptide mapping of this subunit will be used to examine
questions of molecular topography, the location of functionally active
sites, and the location of determinants recognized by monoclonal
antibodies. Quench-flow studies of purified, reconstituted channels will
be extended to further characterize the reconstitution system, the
interrelationship of neurotoxin binding sites in the reconstituted protein,
and the functional integrity of channel protein. Recent work with single
channel measurements on the purified protein will be pursued to measure
kinetic and voltage-dependence properties of the purified reconstituted
channel that are not accessible with the quench-flow system. Ongoing work
with monoclonals will be continued in order to expand our panel of
channel-specific antibodies, and to identify antibodies that interfere with
channel function. Integration of these various approaches is emphasized
with the goal of exploring structure-function relationships in the purified
channel at the molecular level.
StatusFinished
Effective start/end date12/31/8911/30/96

Funding

  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke

ASJC

  • Medicine(all)
  • Neuroscience(all)
  • Immunology

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