Key Points
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The cloning of the complementary DNAs and genes for the insulin and insulin-like growth factor 1 (IGF1) receptors, as well as tertiary-structure predictions, have provided valuable insights into the overall domain organization of the receptors.
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No crystal structure of the insulin- or IGF1-receptor complexes with their ligands is yet available, but crystal structures of the large domain 1 (L1)–Cys-rich (CR)–L2 amino-terminal fragment of the IGF1 receptor, and of the insulin- and IFG1-receptor tyrosine-kinase domains both in the inactive and activated conformation, are available.
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Single-molecule electron-microscopic imaging of the insulin receptor has given some indications of the overall organization of the extracellular receptor domains, although there have been variable results and a tendency in some cases to over-interpret low-resolution data.
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Knowledge of the structure of several receptor tyrosine-kinase domains has led to attempts to screen for, or design, mimetics or inhibitors, with some degree of success. Agonists or antagonists that target the ligand-binding sites might have a greater chance to be selective, hence the importance of understanding the nature of the ligand-binding mechanism.
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Mapping the ligand-binding sites on the insulin and IGF1 receptors by receptor crosslinking with photoreactve ligands, by examining the binding selectivity of chimeric insulin–IGF1 receptors, by alanine-scanning mutagenesis of receptor domains and by reconstitution of minimized receptor constructs with low or high affinity, has provided a wealth of information on the binding epitopes.
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Likewise, mapping of the residues on the insulin molecule that are involved in receptor binding has progressed, and has revealed the existence of a second binding surface in addition to the so-called 'classical' binding surface. The information on IGF1 and IGF2 is more fragmentary.
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Alternative crosslinking models that explain the complex ligand-binding kinetics of the insulin and IGF1 receptors (including negative cooperativity) are discussed.
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Various strategies for designing agonists or antagonists of a dimerizing receptor are discussed, building on the experience acquired with the erythropoietin receptor.
Abstract
Type 2 diabetes mellitus — in which the body produces insufficient amounts of insulin or the insulin that is produced does not function properly to control blood glucose — is an increasingly common disorder. Prospective clinical studies have proven the benefits of tighter glucose control in reducing the frequency and severity of complications of the disease, leading to the advocation of earlier and more aggressive use of insulin therapy. Given the reluctance of patients with type 2 diabetes to inject themselves with insulin, orally active insulin mimetics would be a major therapeutic advance. Here, we discuss recent progress in understanding the structure–function relationships of the insulin and insulin-like growth factor 1 (IGF1) receptors, their mechanism of activation and their implications for the design of insulin-receptor agonists for diabetes therapy and IGF1-receptor antagonists for cancer therapy.
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Acknowledgements
We are grateful to J. Brandt and M. Krogsgaard Thomsen for critical review of the manuscript. The Receptor Biology Laboratory and the Hagedorn Research Institute are independent basic research components of Novo Nordisk A/S. The authors are also supported by grants from the Juvenile Diabetes Research Foundation International, the Danish Research Council through the Danish Center for Growth and Regeneration, and the Øresund region's Research and Development Committee (Øforsk).
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Animated figure 4| Insulin-like growth factor See legend below.
a | The three-dimensional structure of the large domain 1 (L1)–Cys-rich (CR)–L2 domain of the insulin-like growth factor 1 (IGF1) receptor determined by X-ray crystallography 17. An extended bi-lobed structure (40 × 48 × 105 Å) comprises the two globular L-domains with a new type of right-handed β-helix fold that flank the CR domain. They seem to be part of the leucine-rich-repeat superfamily 134. Although L1 (residues 1–150; green) contacts the CR domain (blue) along its length, there is minimal contact with L2 (residues 300–460; orange). The CR domain comprises an array of disulphide-linked modules that resemble those in the tumour-necrosis factor (TNF) receptor and laminin23. The different orientations of L1 and L2 relative to the CR domain are probably an artefact of crystal packing, and the position of L2 is probably more parallel to L1 in the native structure. The flexibility between the CR domain and L2 might be important for ligand binding 16. A cavity of ∼30 Å diameter occupies the centre of the molecule and represents a potential binding pocket, although this construct does not bind IGF1.The amino acids that have been determined by alanine-scanning mutagenesis 80 to be important for ligand binding are shown in yellow as van der Waals spheres (see main text for details). The three-dimensional structure of the IGF1 molecule is shown, based on the X-ray coordinates of Brzozowski and colleagues101. The amino acids that have been determined by site-directed mutagenesis to be important for receptor binding are shown in yellow as van der Waals spheres (reviewed in Ref. 7). As Arg36 and Arg37 are lacking in the structure, residues 35 and 38 have been highlighted instead to show their approximate location in the middle of the C-peptide domain. The backbone is shown in blue. b | The insulin-receptor L1–CR–L2 domain has been modelled on the corresponding domain coordinates of the IGF1-receptor using the programme SwissModel. The amino acids that have been determined by site-directed mutagenesis to be important for receptor binding66 are shown in yellow as van der Waals spheres. The three-dimensional structure of the insulin molecule is shown, based on the X-ray coordinates of Baker and colleagues92. The classical binding surface is shown in yellow as van der Waals spheres; Leu A13 and Leu B17 are shown in red. The backbone is shown in blue. Modelling was carried out using the DS ViewerLite 5.0 (Accelrys). (GIF 321 kb)
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Glossary
- HYPOGLYCAEMIC AGENT
-
A drug that lowers blood glucose.
- TRANS-PHOSPHORYLATION
-
An enzymatic mechanism whereby one receptor-tyrosine-kinase domain transfers the γ-phosphate group of ATP (adenosine trisphosphate) to tyrosyl side chains of the second tyrosine-kinase domain of the same receptor (as opposed to the tyrosine-kinase domain phosphorylating itself, which would be cis-phosphorylation).
- SCATCHARD PLOT
-
A graphical method designed by George Scatchard in 1949 that consists of plotting the ratio of bound/free ligand against the concentration of bound ligand (as in figure 5a). In the case of simple (non-cooperative) binding, the graph is a straight line with a slope of −Ka, the affinity constant of the reaction.
- NEGATIVE COOPERATIVITY
-
A property of a multi-site-binding protein (for example an enzyme or a receptor) whereby the binding of one ligand molecule decreases the binding affinity of other ligand molecules to neighbouring binding sites.
- ALANINE SCANNING
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The systematic replacement of the amino acids of a ligand or a binding protein by alanine (which just has a methyl group side chain) in order to evaluate the importance of the replaced side chains in the binding process.
- HOLORECEPTOR
-
The full-length receptor, as opposed to various minimized constructs, such as those shown in Box 2.
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De Meyts, P., Whittaker, J. Structural biology of insulin and IGF1 receptors: implications for drug design. Nat Rev Drug Discov 1, 769–783 (2002). https://doi.org/10.1038/nrd917
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DOI: https://doi.org/10.1038/nrd917
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