title: Direct force measurements of insulin monomer-monomer interactions.
abstract: Direct measurement of the forces involved in protein-protein and protein-receptor interactions can, in principle, provide insight necessary for the advancement of structural biology, molecular biology, and the development of therapeutic proteins. The protein insulin is illustrative in this respect as the mechanisms of insulin dimer dissociation and insulin-insulin receptor binding are crucial to the efficacy of insulin medications for the control of diabetes. Insulin molecules, modified with a photochemically active azido functionality on specific residues, were attached to force microscope tips and opposing mica surfaces in configurations that would either favor or disfavor dimer formation. Force curve measurements performed in buffer solution revealed the complexity of the insulin monomer-monomer interaction with multiple unbinding events occurring upon tip retraction, suggesting disruption of discrete molecular bonds at the monomer-monomer interface. Furthermore, the force curves exhibit long-range unbinding events, consistent with considerable elongation of the insulin molecule prior to dissociation. The unbinding forces observed in this study are the result of a combination of molecular disentanglement and dimer dissociation processes.
authors.affiliation: Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis 55455, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Yip
authors.forename: C C
authors.initials: CC
authors.lastname: Yip
authors.forename: M D
authors.initials: MD
authors.lastname: Ward
mesh_terms: D000538:Aluminum Silicates; D000595:Amino Acid Sequence; D000818:Animals; D002021:Buffers; D002417:Cattle; D018360:Crystallography, X-Ray; D019281:Dimerization; D006801:Humans; D007328:Insulin / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding; D013499:Surface Properties
chemical_list: D000538:Aluminum Silicates; D002021:Buffers; D007328:Insulin; C011934:mica