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Updated: Mar 21, 2026
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Published on: May 20, 2020
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Functional evolution of IGF2:IGF2R domain 11 binding generates novel structural interactions and a specific IGF2
Susana Frago1, Ryan D Nicholls2, Madeleine Strickland2
1Tumour Growth Control Group, Oxford Molecular Pathology Institute, Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom;
Summary
Researchers engineered a modified mannose 6-phosphate/insulin-like growth factor-2 receptor (M6P/IGF2R) domain 11 to enhance binding affinity for IGF2. This modification resulted in a 100-fold increase in affinity, creating a potential therapeutic for IGF2-driven cancers.
Area of Science:
- Biochemistry
- Structural Biology
- Molecular Medicine
Background:
- The mannose 6-phosphate/insulin-like growth factor-2 receptor (M6P/IGF2R) plays a role in regulating insulin-like growth factor 2 (IGF2) bioavailability.
- Domain 11 of M6P/IGF2R specifically binds IGF2, influencing ligand levels and mammalian growth.
- Previous studies indicated that the IGF2 binding site in domain 11 is amenable to modification for enhanced affinity.
Purpose of the Study:
- To investigate the potential for enhancing the binding affinity and specificity of M6P/IGF2R domain 11 for IGF2.
- To elucidate the structural mechanisms underlying affinity improvements.
- To assess the therapeutic potential of engineered M6P/IGF2R domain 11 variants.
Main Methods:
- Yeast surface display was employed for mutant selection, guided by high-resolution structure-based predictions.
- Surface plasmon resonance was used to validate binding affinities.
- Nuclear Magnetic Resonance (NMR) spectroscopy confirmed structural changes, specifically increased rigidity in the AB loop.
- Combinatorial mutations were introduced into ligand-interacting loops (AB, CD, FG, HI).
Main Results:
- Previously unidentified mutations were discovered in key ligand-interacting loops.
- Combined mutations in the AB loop increased its rigidity, confirmed by NMR.
- The integration of AB loop mutations with CD and FG loop mutations yielded a 100-fold selective increase in IGF2 binding affinity.
- Structural analysis revealed improved shape complementarity due to interloop and intraloop side chain interactions.
- Engineered domain 11 variants functioned as soluble antagonists, depleting serum IGF2 and inhibiting IGF2-dependent signaling in vivo.
Conclusions:
- Re-engineering M6P/IGF2R domain 11 can significantly enhance IGF2 binding affinity and specificity.
- The structural basis for enhanced affinity involves improved loop rigidity and complementarity.
- Engineered M6P/IGF2R domain 11 variants show promise as therapeutic agents for conditions with IGF2 dysregulation, such as certain cancers.
Keywords:
binding kineticsbiological therapygrowth factor receptorinsulin-like growth factor 2protein evolutionMore Related Videos
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