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Direct insight into insulin aggregation by 2D NMR complemented by PFGSE NMR
Author(s) -
Bocian Wojciech,
Sitkowski Jerzy,
Tarnowska Anna,
Bednarek Elżbieta,
Kawȩcki Robert,
Koźmiński Wiktor,
Kozerski Lech
Publication year - 2008
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.21969
Subject(s) - dilution , chemistry , two dimensional nuclear magnetic resonance spectroscopy , insulin , nmr spectra database , zinc , diffusion , equilibrium constant , analytical chemistry (journal) , chromatography , spectral line , thermodynamics , stereochemistry , organic chemistry , physics , biology , astronomy , endocrinology
The aggregation of Zn(II)‐bound and zinc‐free human insulin was studied in solution using the H β ∼CH 3 crosspeaks 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. Proteins 2008. © 2008 Wiley‐Liss, Inc.