Identification of a binding sequence for the 14-3-3 protein within the cytoplasmic domain of the adhesion receptor, platelet glycoprotein Ib alpha

J Biol Chem. 1996 Mar 29;271(13):7362-7. doi: 10.1074/jbc.271.13.7362.

Abstract

The zeta-form 14-3-3 protein (14-3-3zeta) regulates protein kinases and interacts with several signaling molecules. We reported previously that a platelet adhesion receptor, glycoprotein (GP) Ib-IX, was associated with a 29-kDa protein with partial sequences identical to 14-3-3zeta. In this study, the interaction between GPIb-IX and recombinant 14-3-3zeta is reconstituted. Further, we show that the 14-3-3zeta binding site in GPIb is within a 15 residue sequence at the C terminus of GPIb-alpha, as indicated by antibody inhibition and direct binding of 14-3-3zeta to synthetic GPIb-alpha cytoplasmic domain peptides. The 14-3-3zeta binds to recombinant wild type GPIb-IX but not to the GPIb-alpha mutants lacking C-terminal 5 or more residues, suggesting that the C-terminal 5 residues of GPIb-alpha are critical. Similarity between the GPIb-alpha C-terminal sequence and the serine-rich regions of Raf and Bcr kinases suggests a possible serine-rich recognition motif for the 14-3-3 protein.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 14-3-3 Proteins
  • Amino Acid Sequence
  • Animals
  • Antibodies
  • Base Sequence
  • Binding Sites
  • CHO Cells
  • Cell Line
  • Chromatography, Affinity
  • Cloning, Molecular
  • Cricetinae
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Gene Library
  • Humans
  • Immunoblotting
  • Kinetics
  • Molecular Sequence Data
  • Peptides / chemical synthesis
  • Peptides / immunology
  • Platelet Glycoprotein GPIb-IX Complex / chemistry
  • Platelet Glycoprotein GPIb-IX Complex / isolation & purification
  • Platelet Glycoprotein GPIb-IX Complex / metabolism*
  • Polymerase Chain Reaction
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / isolation & purification
  • Proteins / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Serine
  • Signal Transduction
  • Transfection
  • Tumor Cells, Cultured
  • Tyrosine 3-Monooxygenase*

Substances

  • 14-3-3 Proteins
  • Antibodies
  • DNA Primers
  • Peptides
  • Platelet Glycoprotein GPIb-IX Complex
  • Proteins
  • Recombinant Proteins
  • Serine
  • Tyrosine 3-Monooxygenase