Sequence Specificity of C‐Terminal Src Kinase (Csk)
10.1111/j.1432-1033.1997.t01-1-00433.x
Crossref journal-article
Wiley
European Journal of Biochemistry (311)
Abstract

An eicosapeptide encompassing the C‐terminal tail of c‐Src (Tyr527) which is conserved in most Src‐ related protein kinases, is phosphorylated by C‐terminal Src kinase (CSK) and by the two Src‐related protein kinases c‐Fgr and Lyn, with similar kinetic constants. Two related peptides reproducing the C‐terminal segments of c‐Src mutants defective in CSK phosphorylation [MacAuley, A., Okada, M., Nada, S., Nakagawa, H. & Cooper, J. A. (1993) Oncogene 8, 117–124] AFLEDSCTGTEPLYQRGENL (mutant number 28) and AFLEDNFTGTKPQYHPGENL (mutant number 29), proved a better and a much worse substrates, respectively than the wild‐type peptide, with either CSK or the two Src kinases. By changing individual residues in the best peptide substrate, it was shown that the main element responsible for its improved phosphorylation is leucine at position –1 (instead of glutamine), while lysine at position –3 (instead of glutamate) has a detrimental effect, possibly accounting for the negligible phosphorylation of peptide derived from mutant number 29. By contrast to most peptide substrates, including the Src C‐terminal peptides, which exhibit relatively high Km values, a polyoma‐virus‐middle‐T‐antigen‐(mT)‐derived peptide with tyrosine embedded in a highly hydrophobic sequence (EEEPQFEEIPIYLELLP) exhibits with CSK a quite low Km value (63 μM). Consistent with this, the optimal sequence selected by CSK in an oriented peptide library is XXXIYMFFF. This is different from sequences selected by Lyn (DEEIYEELX) and c‐Fgr (XEEIYGIFF), although they all share a high selection for a hydrophobic residue at n–1. In sharp contrast, TPKIIB/p38Syk, related to the catalytic domain of p72syk, selects acidic residues at nearly all positions, n–1 included.These data support the notion that the features determining the specific phosphorylation of the C‐terminal tyrosine residue of Src do not reside in the primary structure surrounding the target tyrosine. They also show that this site does not entirely fulfil the optimal consensus sequence recognized by CSK, disclosing the possibility that as yet unrecognized CSK targets structurally unrelated to the C‐terminal tyrosine residue of Src kinases may exist.

Bibliography

Ruzzene, M., Songyang, Z., Marin, O., Donella‐Deana, A., Brunati, A. M., Guerra, B., Agostinis, P., Cantley, L. C., & Pinna, L. A. (1997). Sequence Specificity of C‐Terminal Src Kinase (Csk). European Journal of Biochemistry, 246(2), 433–439. Portico.

Authors 9
  1. Maria Ruzzene (first)
  2. Zhou Songyang (additional)
  3. Oriano Marin (additional)
  4. Arianna Donella‐Deana (additional)
  5. Anna Maria Brunati (additional)
  6. Barbara Guerra (additional)
  7. Patrizia Agostinis (additional)
  8. Lewis C. Cantley (additional)
  9. Lorenzo A. Pinna (additional)
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Dates
Type When
Created 21 years, 1 month ago (July 16, 2004, 8:31 a.m.)
Deposited 1 year, 10 months ago (Oct. 16, 2023, 12:57 p.m.)
Indexed 1 year, 10 months ago (Oct. 16, 2023, 1:10 p.m.)
Issued 28 years, 3 months ago (June 1, 1997)
Published 28 years, 3 months ago (June 1, 1997)
Published Online 21 years, 1 month ago (July 16, 2004)
Published Print 28 years, 3 months ago (June 1, 1997)
Funders 0

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@article{Ruzzene_1997, title={Sequence Specificity of C‐Terminal Src Kinase (Csk): A Comparison with Src‐Related Kinases C‐Fgr and Lyn}, volume={246}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1997.t01-1-00433.x}, DOI={10.1111/j.1432-1033.1997.t01-1-00433.x}, number={2}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Ruzzene, Maria and Songyang, Zhou and Marin, Oriano and Donella‐Deana, Arianna and Brunati, Anna Maria and Guerra, Barbara and Agostinis, Patrizia and Cantley, Lewis C. and Pinna, Lorenzo A.}, year={1997}, month=jun, pages={433–439} }