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Structural insights into the interaction of a monoclonal antibody and Nodal peptides by STD-NMR spectroscopy
Luisa Calvanese1, Annalia Focà2, Annamaria Sandomenico3
1CIRPeB, University of Naples Federico II, via Mezzocannone, 16, 80134 Napoli, Italy.
Abstract:
Nodal is a growth factor expressed during early embryonic development, but reactivated in several advanced-stage cancers. Targeting of Nodal signaling, which occurs via the binding to Cripto-1 co-receptor, results in inhibition of cell aggressiveness and reduced tumor growth. The Nodal binding region to Cripto-1 was identified and targeted with a high affinity monoclonal antibody (3D1). By STD-NMR technique, we investigated the interaction of Nodal fragments with 3D1 with the aim to elucidate at atomic level the interaction surface. Data indicate with high accuracy the antibody-antigen contact atoms and confirm the information previously obtained by immune-enzymatic methods. Main residues contacted by 3D1 are P46, V47, E49 and E50, which belong to the Nodal loop involved in the interaction with the co-receptor.
Insights
Researchers mapped the atomic-level interactions between the Nodal growth factor and a high-affinity antibody (3D1). This study elucidates critical binding sites, aiding in the development of targeted cancer therapies by inhibiting Nodal signaling.
Area of Science:
- Molecular Biology
- Structural Biology
- Oncology
Background:
- Nodal is a growth factor crucial in embryonic development, often reactivated in advanced cancers.
- Nodal signaling, mediated by the Cripto-1 co-receptor, promotes cancer cell aggressiveness and tumor growth.
- Targeting Nodal signaling offers a therapeutic strategy to inhibit cancer progression.
Purpose of the Study:
- To identify and characterize the atomic-level interaction surface between Nodal and its co-receptor binding region.
- To investigate the binding interface of Nodal with the high-affinity monoclonal antibody (3D1) using biophysical techniques.
- To confirm and refine previous findings on Nodal-Cripto-1 interactions at the molecular level.
Main Methods:
- Saturation Transfer Difference Nuclear Magnetic Resonance (STD-NMR) spectroscopy was employed to study molecular interactions.
- Analysis of Nodal fragments interacting with the 3D1 antibody.
- Comparison of NMR data with previous immune-enzymatic assay results.
Main Results:
- The study precisely mapped the antibody-antigen contact points at an atomic level.
- Key residues P46, V47, E49, and E50 on Nodal were identified as primary contact sites for the 3D1 antibody.
- These residues are located within the Nodal loop critical for Cripto-1 co-receptor binding.
Conclusions:
- The 3D1 antibody binds to a specific Nodal surface involved in co-receptor interaction.
- Structural insights at the atomic level validate the antibody's binding epitope.
- This detailed understanding facilitates the rational design of Nodal-targeting therapeutics for cancer treatment.
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