CONFORMATIONS OF INSULIN &ITS RECEPTOR COMPLEXES

Project: Research project

Project Details

Description

The relationship of structure and bioactivity in insulin is
complex. Determination of a solution conformation may yield
insight into its activity, and establish methods for studying
insulin's interaction with its natural receptors, and rational
design of new drugs for diabetes mellitus. It is proposed to study
the conformations of several derivatives of differing activity,
synthetic analogs similar to known mutant insulins in humans, and
new analogs designed to test possible structure/activity relations.
The major approach will be application of Nuclear Magnetic
Resonance (NMR) techniques to insulin analogs specifically designed
and synthesized for those studies. We shall assign spectra and
detect protein secondary structure by a combination of conventional
and two-dimensional homo- and heteronuclear techniques. Distance
geometry calculations will be used to produce structural
information on those portions of the analogs which are
conformationally static, from nuclear Overhauser measurements
(nOe). Comparative conformational analyses of the analogs will
test the possible relation of the solution structure to
bioactivity. Molecular dynamics will be used to model flexible
regions of the hormone. With the long term view of extending these correlations to direct
measurements of the hormone/receptor complex, we shall apply new
methods of NMR, for connectivity and nOe measurements, which use
detection of protons, and secondary irradiation of 13C and 15N at
labeled sites in insulin analogs. The objectives of these methods,
are the selection of proton homonuclear connectivity and nOe data
for atoms spincoupled to 13C or 15N, and the indirect measurement
of 13C and 15N NMR values in, hormone-receptor complexes. We shall
attempt to identify fragments of insulin receptor suitable for
complex formation and study such complexes. The conformational information derived will be used to design
possible analogs which might test the role of flexibility and
orientation of various parts of insulin. These analogs will be
synthesized, assayed, and conformationally characterized, by NMR
and as time permits, directly in complexes with receptor fragments.
StatusFinished
Effective start/end date8/1/778/31/93

Funding

  • National Institutes of Health

ASJC

  • Medicine(all)
  • Drug Discovery
  • Structural Biology
  • Biophysics
  • Biochemistry
  • Organic Chemistry

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