Interaction of insulin-like growth factor binding protein-4, Miz-1, leptin, lipocalin-type prostaglandin D synthase, and granulin precursor with the N-terminal half of type III hexokinase

Arch Biochem Biophys. 2000 Oct 15;382(2):262-74. doi: 10.1006/abbi.2000.2019.

Abstract

Insulin-like growth factor binding protein-4, Miz-1, leptin, prostaglandin D synthase, and granulin precursor were identified as proteins interacting with the N-terminal half of mammalian Type III hexokinase (HKIII) in the yeast two-hybrid method. These interactions were confirmed by in vitro binding studies. All five of these proteins, and their mRNAs, were present in PC12 cells, as shown by immunoblotting and RT-PCR, respectively. All were coimmunoprecipitated from PC12 extracts with an antibody against HKIII, but not with anti-Type I hexokinase. Moreover, all of these proteins were coimmunoprecipitated using antileptin as precipitating antibody, indicating the existence of a macromolecular complex including these five proteins and HKIII. Transfection of M+R 42 cells with HKIII-green fluorescent protein (GFP) reporter constructs gave a diffuse intracellular fluorescence. Cotransfection with leptin or Miz-1 resulted in distinctly different localization of the HKIII-GFP fusion protein, at intracellular sites coincident with localization of leptin-GFP or Miz-1-GFP reporter constructs.

Publication types

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

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • DNA Primers / genetics
  • DNA-Binding Proteins / metabolism
  • Glycoproteins / metabolism
  • Green Fluorescent Proteins
  • Growth Substances / metabolism
  • Hexokinase / chemistry*
  • Hexokinase / genetics
  • Hexokinase / metabolism*
  • In Vitro Techniques
  • Insulin-Like Growth Factor Binding Protein 4 / metabolism
  • Intercellular Signaling Peptides and Proteins*
  • Intramolecular Oxidoreductases / metabolism
  • Leptin / metabolism
  • Lipocalins
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • PC12 Cells
  • Progranulins
  • Protein Binding
  • Rats
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Transfection
  • Two-Hybrid System Techniques

Substances

  • DNA Primers
  • DNA-Binding Proteins
  • Glycoproteins
  • Growth Substances
  • Insulin-Like Growth Factor Binding Protein 4
  • Intercellular Signaling Peptides and Proteins
  • Leptin
  • Lipocalins
  • Luminescent Proteins
  • Progranulins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Hexokinase
  • Intramolecular Oxidoreductases
  • prostaglandin R2 D-isomerase