Direct determination of the insulin-insulin receptor interface using transferred cross-saturation experiments.
Insulin initiates metabolic control by binding to the insulin receptor (IR) on target cells. Kinetic and mutational analyses have revealed two binding sites on the insulin molecule and the residues that compose them. However, direct determination of the insulin-IR interface is required to distinguish those residues that contribute to receptor binding from those required for structural stability. Here, we successfully characterized one binding site using the nuclear magnetic resonance (NMR) transferred cross-saturation method, which can directly determine the binding interface of a large protein-protein complex. The results showed that this binding site contained three residues that have not been identified previously by mutational analyses. On the basis of the structure of the contact site, we also identified a molecule that can displace insulin from the IR. In addition, we discuss the mode of interaction between insulin and its receptor relative to the NMR analyses.
The Institute of Life Sciences, Ajinomoto Co Inc, 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
Takefumi
T
Nakamura
Hideo
H
Takahashi
Mitsuo
M
Takahashi
Nobuhisa
N
Shimba
Ei-ichiro
E
Suzuki
Ichio
I
Shimada
D000595:Amino Acid Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D007328:Insulin / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019906:Nuclear Magnetic Resonance, Biomolecular; D011972:Receptor, Insulin / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
D007328:Insulin; D011972:Receptor, Insulin