Journal
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
Volume 71, Issue 3, Pages 1057-1065Publisher
WILEY
DOI: 10.1002/prot.21969
Keywords
human insulin aggregates; diffusion coefficients; PFGSE NMR; oligomer distribution; association constants
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The aggregation of Zn(II)bound and zinc-free human insulin was studied in solution using the H-beta similar to CH3 cross-peaks of threonine residues in 2D COSY, TOCSY, and NOESY NMR spectra which allow viewing of the oligomers in equilibrium. This is complemented by PFGSE measurements of the translational diffusion coefficient, D-i, used for monitoring the changes in equilibrium composition of aggregates on dilution of both insulins in physiological medium. The back calculation of the dilution isotherm allows establishing the association constants for oligomeric equilibria in solution and discussion of the models of association.
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