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A novel, specific interaction involving the Csk SH3 domain and its natural ligand

Nature Structural Biology volume 8, pages 998–1004 (2001)Cite this article

Abstract

C-terminal Src kinase (Csk) takes part in a highly specific, high affinity interaction via its Src homology 3 (SH3) domain with the proline-enriched tyrosine phosphatase PEP in hematopoietic cells. The solution structure of the Csk-SH3 domain in complex with a 25-residue peptide from the Pro/Glu/Ser/Thr-rich (PEST) domain of PEP reveals the basis for this specific peptide recognition motif involving an SH3 domain. Three residues, Ala 40, Thr 42 and Lys 43, in the SH3 domain of Csk specifically recognize two hydrophobic residues, Ile 625 and Val 626, in the proline-rich sequence of the PEST domain of PEP. These two residues are C-terminal to the conventional proline-rich SH3 domain recognition sequence of PEP. This interaction is required in addition to the classic polyproline helix (PPII) recognition by the Csk-SH3 domain for the association between Csk and PEP in vivo. NMR relaxation analysis suggests that Csk-SH3 has different dynamic properties in the various subsites important for peptide recognition.

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Figure 1: Sequence alignment for several SH3 domains involved with Pro-rich peptide binding and Pro-rich peptides from tyrosine phosphatases, PEP and PTP-PEST.
Figure 2: Solution structure of the Csk-SH3–PEP-3BP1 complex.
Figure 3: Detailed view of the two distinct surfaces on Csk-SH3 involved in peptide recognition.
Figure 4: Backbone dynamics of the Csk-SH3–PEP-3BP1 complex determined by 15N relaxation measurements.
Figure 5: Comparison of the peptide recognition surfaces in the HIV-1 Nef–Fyn-SH3 (R96I)33 mutant and Csk-SH3–PEP interactions.

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Acknowledgements

R.G. would like to thank D. Fushman for useful discussions and for providing the DYNAMICS package and P. Loria for providing the relaxation compensated CPMG pulse sequence. The authors thank P.A. Cole for useful discussions. This work has been supported by a grant from the National Institutes of Health and a fellowship from the National Cancer Institute to A.S.

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  1. Ranajeet Ghose, Alexander Shekhtman and David Cowburn: These authors contributed equally to this work.

Authors and Affiliations

  1. The Rockefeller University, 1230 York Avenue, New York, 10021-6399, New York, USA

    Ranajeet Ghose, Alexander Shekhtman, Michael J. Goger, Hong Ji & David Cowburn

  2. The New York Structural Biology Center, c/o Box 163, 1230 York Avenue, New York, 10021-6399, New York, USA

    David Cowburn

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Correspondence to David Cowburn.

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Ghose, R., Shekhtman, A., Goger, M. et al. A novel, specific interaction involving the Csk SH3 domain and its natural ligand. Nat Struct Mol Biol 8, 998–1004 (2001). https://doi.org/10.1038/nsb1101-998

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