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High-performance liquid chromatography of insulin. Accessibility and flexibility
A N McLeod1, A Auf der Mauer, S P Wood
1Department of Crystallography, Birkbeck College, University of London, U.K.
Journal of Chromatography
|March 2, 1990
Summary
Insulin monomer conformation changes are key to receptor interaction. Analytical chromatography revealed partial unfolding of insulin monomers, supporting theories of its biologically active structure.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Insulin receptor interaction is crucial for glucose regulation.
- Conformational changes in the insulin monomer are hypothesized to mediate receptor binding.
Purpose of the Study:
- To investigate the solution conformation of the insulin monomer.
- To correlate structural changes with insulin's biological activity.
Main Methods:
- Analytical reversed-phase high-performance liquid chromatography (RP-HPLC) of insulin analogues.
- Calculation of surface accessibilities for individual amino acid residues.
- Interpretation of elution coefficients based on residue accessibility.
Main Results:
- Evidence for partial unfolding of the insulin monomer under experimental conditions.
- Elution behavior correlated with calculated surface accessibility of residues.
- Data supports a dynamic structural model for insulin.
Conclusions:
- The study provides experimental support for the role of insulin monomer conformational changes in receptor interaction.
- Partial unfolding may represent a biologically relevant conformation of insulin.
- Analytical RP-HPLC is a valuable tool for probing protein solution structures.