Alanine scanning of a putative receptor binding surface of insulin-like growth factor-I
Open accessAbstract
Current evidence supports a binding model in which the insulin molecule contains two binding surfaces, site 1 and site 2, which contact the two halves of the insulin receptor. The interaction of these two surfaces with the insulin receptor results in a high affinity cross-linking of the two receptor alpha subunits and leads to receptor activation. Evidence suggests that insulin-like growth factor-I (IGF-I) may activate the IGF-I receptor in a similar mode. So far IGF-I residues structurally corresponding to the residues of the insulin site 1 together with residues in the C-domain of IGF-I have been found to be important for binding of IGF-I to the IGF-I receptor (e.g. Phe(23), Tyr(24), Tyr(31), Arg(36), Arg(37), Val(44), Tyr(60), and Ala(62)). However, an IGF-I second binding surface similar to site 2 of insulin has not been identified yet. In this study, we have analyzed whether IGF-I residues corresponding to the six residues of the insulin site 2 have a role in high affinity binding of IGF-I to the IGF-I receptor. Six single-substituted IGF-I analogues were produced, each containing an alanine substitution in one of the following positions (corresponding insulin residues in parentheses): Glu(9) (His(B10)), Asp(12) (Glu(B13)), Phe(16) (Leu(B17)), Asp(53) (Ser(A12)), Leu(54) (Leu(A13)), and Glu(58) (Glu(A17)). In addition, two analogues with 2 and 3 combined alanine substitutions were also produced (E9A,D12A IGF-I and E9A,D12A,E58A IGF-I). The results show that introducing alanine in positions Glu(9), Asp(12), Phe(16), Leu(54), and Glu(58) results in a significant reduction in IGF-I receptor binding affinity, whereas alanine substitution at position 53 had no effect on IGF-I receptor binding. The multiple substitutions resulted in a 33-100-fold reduction in IGF-I receptor binding affinity. These data suggest that IGF-I, in addition to the C-domain, uses surfaces similar to those of insulin in contacting its cognate receptor, although the relative contribution of the side chains of homologous residues varies.
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Human IGF-1 acts via zebrafish receptors because the mature peptide shares about 81% identity with zebrafish IGF-1, including key receptor-binding residues, and human and zebrafish IGF-IR share over 80% similarity with conserved ligand-binding residues.
“We suspected human IGF-1 would act via zebrafish receptors because the mature peptide shares ~81% identity with that of zebrafish (Zou et al., 2009), including key residues important for receptor binding (Gauguin et al., 2008). Furthermore, human and zebrafish IGF-IR share over 80% similarity and key residues involved in ligand binding are preserved (Ayaso et al., 2002; Renteria et al., 2008).”
From Retinal injury, growth factors, and cytokines converge on β-catenin and pStat3 signaling to stimulate retina regeneration (2014) · cites this paper as Gauguin et al., 2008
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References 56 · Cited by 67
Research works printed but not linked: 2
- Lou, M., Garrett, T. P., McKern, N. M., Hoyne, P. A., Epa, V. C., Bentley, J. D., Lovrecz, G. O., Cosgrove, L. J., Frenkel, M. J., and Ward, C. W. (2006) Proc. Natl. Acad. Sci. U. S. A. 103, 12429 -12434
- Kjeldsen, T., Andersen, A. S., Wiberg, F. C., Rasmussen, J. S., Scha ¨ffer, L., Balschmidt, P., Møller, K. B., and Møller, N. P. (1991) Proc. Natl. Acad. Sci.
- Authors
- Gauguin, L., Delaine, C., Alvino, C. L., McNeil, K. A., Wallace, J. C., Forbes, B. E., & De Meyts, P.
- Affiliations
- The University of Adelaide (AU)
- Corresponding author
- Gauguin, Lisbeth
- ORCID
- Forbes, Briony E.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2008-07
- Volume
- 283
- Issue
- 30
- Pages
- 20821-20829
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC3258947
- OpenAlex
- W1971940467
- Cited by
- 67
- References
- 56
- Citations by year
- 2026: 1 · 2025: 4 · 2023: 6 · 2022: 5 · 2021: 1 · 2020: 4 · 2019: 2 · 2018: 4 · 2017: 3 · 2016: 4 · 2015: 3 · 2014: 4 · 2013: 5 · 2012: 3
- FWCI
- 4.34
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Adelaide Research & Scholarship (AR&S) (University of Adelaide) (submittedVersion)
- Funders
- Novo Nordisk
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Growth Hormone and Insulin-like Growth Factors
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Keywords
- insulin-like growth factor I, IGF-1 receptor, receptor binding affinity, alanine scanning, site-directed mutagenesis, receptor activation, C-domain, ligand-receptor interactions, tyrosine
- MeSH terms
- Alanine, Amino Acid Sequence, Animals, Binding Sites, Humans, Insulin, Ligands, Models, Biological, Molecular Sequence Data, Muscles, Protein Binding, Sequence Homology, Amino Acid, Receptor, IGF Type 1, Mice, Rats
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