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Multimeric and differential binding of CIN 85/ CD 2 AP with two atypical proline‐rich sequences from CD 2 and Cbl‐b*
Author(s) -
Ceregido M. Angeles,
GarciaPino Abel,
OrtegaRoldan Jose L.,
Casares Salvador,
López Mayorga Obdulio,
Bravo Jeronimo,
Nuland Nico A. J.,
Azuaga Ana I.
Publication year - 2013
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.12333
Subject(s) - isothermal titration calorimetry , chemistry , protein–protein interaction , sh3 domain , signal transducing adaptor protein , proto oncogene tyrosine protein kinase src , plasma protein binding , stereochemistry , signal transduction , biochemistry
The CD2AP (CD2‐associated protein) and CIN85 (Cbl‐interacting protein of 85 kD a) 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 CD 2 (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 CD 2 AP ‐ SH 3A 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 abstractCIN85 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 )