title: Direct insight into insulin aggregation by 2D NMR complemented by PFGSE NMR.
abstract: The aggregation of Zn(II)-bound and zinc-free human insulin was studied in solution using the H(beta)-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.
authors.affiliation: National Medicines Institute, Warsaw, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Bocian
authors.forename: Jerzy
authors.initials: J
authors.lastname: Sitkowski
authors.forename: Anna
authors.initials: A
authors.lastname: Tarnowska
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Bednarek
authors.forename: Robert
authors.initials: R
authors.lastname: Kawecki
authors.forename: Wiktor
authors.initials: W
authors.lastname: Koźmiński
authors.forename: Lech
authors.initials: L
authors.lastname: Kozerski
mesh_terms: D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D006801:Humans; D007328:Insulin / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D008956:Models, Chemical; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011485:Protein Binding; D015032:Zinc / Q000032:analysis / Q000737:chemistry / Q000378:metabolism
chemical_list: D007328:Insulin; D015032:Zinc