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NMR spectroscopy of 14-3-3zeta reveals a flexible C-terminal extension: differentiation of the chaperone and phosphoserine-binding activities of 14-3-3zeta

Journal Article


Abstract


  • Intracellular 14-3-3 proteins bind to many proteins, via a specific phosphoserine motif, regulating diverse cellular tasks including cell signalling and disease progression. The 14-3-3ζ isoform is a molecular chaperone, preventing the stressinduced aggregation of target proteins in a manner comparable with that of the unrelated sHsps (small heat-shock proteins). 1H-NMR spectroscopy revealed the presence of a flexible and unstructured C-terminal extension, 12 amino acids in length, which protrudes from the domain core of 14-3-3ζ and is similar in structure and length to the C-terminal extension of mammalian sHsps. The extension stabilizes 14-3-3ζ, but has no direct role in chaperone action. Lys 49 is an important functional residue within the ligand-binding groove of 14-3-3ζ with K49E 14-3-3ζ exhibiting markedly reduced binding to phosphorylated and non-phosphorylated ligands. The R18 peptide binds to the binding groove of 14-3-3ζ with high affinity and also reduces the interaction of 14-3-3ζ ligands. However, neither the K49E mutation nor the presence of the R18 peptide affected the chaperone activity of 14-3-3ζ, implying that the C-terminal extension and binding groove of 14-3-3ζ do not mediate interaction with target proteins during chaperone action. Other region(s) in 14-3-3ζ are most likely to be involved, i.e. the protein's chaperone and phosphoserine-binding activities are functionally and structurally separated. © The Authors Journal compilation © 2011 Biochemical Society.

Authors


  •   Williams, Danielle M. (external author)
  •   Ecroyd, Heath
  •   Goodwin, K L. (external author)
  •   Dai, Huanqin (external author)
  •   Fu, H (external author)
  •   Woodcock, Joanna M. (external author)
  •   Zhang, Lixin (external author)
  •   Carver, John A. (external author)

Publication Date


  • 2011

Citation


  • Williams, D. M., Ecroyd, H., Goodwin, K. L., Dai, H., Fu, H., Woodcock, J. M., Zhang, L. & Carver, J. A. (2011). NMR spectroscopy of 14-3-3zeta reveals a flexible C-terminal extension: differentiation of the chaperone and phosphoserine-binding activities of 14-3-3zeta. Biochemical Journal, 437 (3), 493-503.

Scopus Eid


  • 2-s2.0-79960274257

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/3449

Has Global Citation Frequency


Number Of Pages


  • 10

Start Page


  • 493

End Page


  • 503

Volume


  • 437

Issue


  • 3

Abstract


  • Intracellular 14-3-3 proteins bind to many proteins, via a specific phosphoserine motif, regulating diverse cellular tasks including cell signalling and disease progression. The 14-3-3ζ isoform is a molecular chaperone, preventing the stressinduced aggregation of target proteins in a manner comparable with that of the unrelated sHsps (small heat-shock proteins). 1H-NMR spectroscopy revealed the presence of a flexible and unstructured C-terminal extension, 12 amino acids in length, which protrudes from the domain core of 14-3-3ζ and is similar in structure and length to the C-terminal extension of mammalian sHsps. The extension stabilizes 14-3-3ζ, but has no direct role in chaperone action. Lys 49 is an important functional residue within the ligand-binding groove of 14-3-3ζ with K49E 14-3-3ζ exhibiting markedly reduced binding to phosphorylated and non-phosphorylated ligands. The R18 peptide binds to the binding groove of 14-3-3ζ with high affinity and also reduces the interaction of 14-3-3ζ ligands. However, neither the K49E mutation nor the presence of the R18 peptide affected the chaperone activity of 14-3-3ζ, implying that the C-terminal extension and binding groove of 14-3-3ζ do not mediate interaction with target proteins during chaperone action. Other region(s) in 14-3-3ζ are most likely to be involved, i.e. the protein's chaperone and phosphoserine-binding activities are functionally and structurally separated. © The Authors Journal compilation © 2011 Biochemical Society.

Authors


  •   Williams, Danielle M. (external author)
  •   Ecroyd, Heath
  •   Goodwin, K L. (external author)
  •   Dai, Huanqin (external author)
  •   Fu, H (external author)
  •   Woodcock, Joanna M. (external author)
  •   Zhang, Lixin (external author)
  •   Carver, John A. (external author)

Publication Date


  • 2011

Citation


  • Williams, D. M., Ecroyd, H., Goodwin, K. L., Dai, H., Fu, H., Woodcock, J. M., Zhang, L. & Carver, J. A. (2011). NMR spectroscopy of 14-3-3zeta reveals a flexible C-terminal extension: differentiation of the chaperone and phosphoserine-binding activities of 14-3-3zeta. Biochemical Journal, 437 (3), 493-503.

Scopus Eid


  • 2-s2.0-79960274257

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/3449

Has Global Citation Frequency


Number Of Pages


  • 10

Start Page


  • 493

End Page


  • 503

Volume


  • 437

Issue


  • 3