STRUCTURE/FUNCTION ANALYSIS OF ACCHR EPITOPES

Project: Research project

Description

The central theme of this Program Project is to use monoclonal
antibodies (mAbs) directed against defined oligopeptides within
the subunits of the nicotinic acetylcholine receptor (AcChR) as
site-specific probes of AcChR structure (both primary and
tertiary structure) and AcChR function. Special attention will be
paid to mAbs whose binding interferes with the function of this
transmembrane glycoprotein neurotransmitter receptor. Projects
1-4 will assess the effects of the mAbs on particular aspects of
AcChR function: Project 1 - ligand binding, Project 2 - ion fluxes
by stopped flow technique; Project 3 - ion channel function by
patch clamp technique; Project 4 - neuromuscular transmission in
vitro and in vivo. Projects 5 and 6 will localize the epitopes of
these mAbs within the AcChR structure: Project 5 - production
of rationally chosen synthetic oligopeptides mimicking specific
sites within AcChR subunits, assessment of the contribution of
glycosyl moieties to mAb epitopes, Project 6 -identification of
bound mAbs on the low resolution three-dimensional model of
AcChR by both small angle X-ray diffraction and high resolution
electron microscopy, as well as localization of fluorescent probes.
The Scientific Core will produce new mAbs directed against the
peptides produced in Project 5 and will provide purified,
characterized mAbs for the various projects. In Project 4, the
peptides will be used to possibly develop a new treatment
modality for experimental myasthenia gravis. This Program
Project represents a collaborative effort of immunologists,
biochemists, electrophysiologists, peptide chemists, and
physical/structural chemists in which the information from the
various projects, taken together, will allow for mapping of various
sites in the primary sequence of AcChR, especially functionally
important sites, onto the three-dimensional model.
StatusFinished
Effective start/end date1/1/879/30/92

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $180,591.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

Fingerprint

Cholinergic Receptors
Epitopes
acetylcholine
Blocking Antibodies
Monoclonal Antibodies
Ions
Bungarotoxins
Autoimmune Experimental Myasthenia Gravis
three dimensional models
Peptides
Electron microscopy
neuromuscular transmission
Antibodies
Neuromuscular Junction
Myasthenia Gravis
Electromyography
Coloring Agents
Libraries
X ray diffraction
Histology

ASJC

  • Medicine(all)
  • Neuroscience(all)