Multimeric and differential binding of CIN85/CD2AP with two atypical proline‐rich sequences from CD2 and Cbl‐b

• Published in: The FEBS journal Vol. 280; no. 14; pp. 3399 - 3415
• Main Authors: Ceregido, M. Angeles, Garcia‐Pino, Abel, Ortega‐Roldan, Jose L, Casares, Salvador, López Mayorga, Obdulio, Bravo, Jeronimo, Nuland, Nico A. J, Azuaga, Ana I
• Format: Journal Article
• Language: English
• Published: England Blackwell 01.07.2013; Blackwell Publishing Ltd
• Subjects: Adaptor Proteins, Signal Transducing - chemistry; Amino Acid Sequence; calorimetry; CD2 Antigens - chemistry; cell adhesion; Cell adhesion & migration; CIN85/CD2AP; crystal structure; Cytoskeletal Proteins - chemistry; Humans; Hydrogen Bonding; ITC; Kinases; ligands; lymphoma; Models, Molecular; Molecular Sequence Data; NMR; nuclear magnetic resonance spectroscopy; Nuclear Magnetic Resonance, Biomolecular; Proline; Protein Binding; Protein Structure, Secondary; Proteins; Proto-Oncogene Proteins c-cbl - chemistry; receptors; SAXS; Scattering, Small Angle; SH3 domain; Signal transduction; src Homology Domains; T cell receptors; T-lymphocytes; Thermodynamics; titration; Titrimetry; X-radiation; X-Ray Diffraction; SH3 domain; ITC; NMR; CIN85/CD2AP; SAXS
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/febs.12333

The CD2AP (CD2‐associated protein) and CIN85 (Cbl‐interacting protein of 85 kDa) adaptor proteins each employ three Src homology 3 (SH3) domains to cluster protein partners and ensure efficient signal transduction and down‐regulation of tyrosine kinase receptors. Using NMR, isothermal titration calorimetry and small‐angle X‐ray scattering methods, we have characterized several binding modes of the N‐terminal SH3 domain (SH3A) of CD2AP and CIN85 with two natural atypical proline‐rich regions in CD2 (cluster of differentiation 2) and Cbl‐b (Casitas B‐lineage lymphoma), and compared these data with previous studies and published crystal structures. Our experiments show that the CD2AP‐SH3A domain forms a type II dimer with CD2 and both type I and type II dimeric complexes with Cbl‐b. Like CD2AP, the CIN85‐SH3A domain forms a type II complex with CD2, but a trimeric complex with Cbl‐b, whereby the type I and II interactions take place at the same time. Together, these results explain how multiple interactions among similar SH3 domains and ligands produce a high degree of diversity in tyrosine kinase, cell adhesion or T‐cell signaling pathways. STRUCTURED DIGITAL ABSTRACT: CIN85 and Cbl-b bind by nuclear magnetic resonance (View Interaction: 1, 2) CD2 and CIN85 bind by nuclear magnetic resonance (View interaction) CD2 and CD2AP bind by nuclear magnetic resonance (View interaction) Cbl-b and CD2AP bind by nuclear magnetic resonance (View Interaction: 1, 2) Cbl-b and CIN85 bind by isothermal titration calorimetry (View Interaction: 1, 2, 3, 4) CIN85 and Cbl-b bind by X-ray scattering (View Interaction: 1, 2) CIN85 and CD2 bind by isothermal titration calorimetry (View interaction) CD2 and CD2AP bind by X-ray scattering (View interaction) CD2AP and CD2 bind by isothermal titration calorimetry (View interaction) CD2 and CIN85 bind by X-ray scattering (View interaction) CD2AP and Cbl-b bind by isothermal titration calorimetry (View Interaction: 1, 2, 3, 4, 5)