1. Academic Validation
  2. Isolation of high-affinity peptide antagonists of 14-3-3 proteins by phage display

Isolation of high-affinity peptide antagonists of 14-3-3 proteins by phage display

  • Biochemistry. 1999 Sep 21;38(38):12499-504. doi: 10.1021/bi991353h.
B Wang 1 H Yang Y C Liu T Jelinek L Zhang E Ruoslahti H Fu
Affiliations

Affiliation

  • 1 Rammelkamp Center for Research, Case Western Reserve University, Cleveland, Ohio 44109, USA.
Abstract

The 14-3-3 proteins interact with diverse cellular molecules involved in various signal transduction pathways controlling cell proliferation, transformation, and Apoptosis. To aid our investigation of the biological function of 14-3-3 proteins, we have set out to identify high-affinity antagonists. By screening phage display libraries, we have identified a set of Peptides which bind 14-3-3 proteins. One of these Peptides, termed R18, exhibited a high affinity for different isoforms of 14-3-3 with estimated K(D) values of 7-9 x 10(-)(8) M. Recognition of multiple isoforms of 14-3-3 suggests the targeting of R18 to a structure that is common among 14-3-3 proteins, such as the conserved ligand-binding groove. Indeed, mutations that alter critical residues in the ligand-binding site of 14-3-3 drastically decreased the level of 14-3-3-R18 association. R18 efficiently blocked the binding of 14-3-3 to the kinase Raf-1, a physiological ligand of 14-3-3, and effectively abolished the protective role of 14-3-3 against phosphatase-induced inactivation of Raf-1. The cocrystal structure of R18 in complex with 14-3-3zeta revealed the occupancy of the general binding groove of 14-3-3zeta by R18, explaining the potent inhibitory effect of R18 on 14-3-3-ligand interactions. Such a well-defined peptide will be an effective tool for probing the role of 14-3-3 in various signaling pathways, and may lead to the development of 14-3-3 antagonists with pharmacological applications.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-P1039A
    99.91%, 14-3-3 Peptide Antagonist
    Raf
  • HY-P1039
    Peptide Antagonists
    Raf