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.
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.
Sui
D
D
Department of Biochemistry, Michigan State University, East Lansing 48824, USA.
Wilson
J E
JE
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006133:Growth Substances / Q000378:metabolism; D006593:Hexokinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D066298:In Vitro Techniques; D018974:Insulin-Like Growth Factor Binding Protein 4 / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D019746:Intramolecular Oxidoreductases / Q000378:metabolism; D020738:Leptin / Q000378:metabolism; D054834:Lipocalins; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016716:PC12 Cells; D000077153:Progranulins; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014162:Transfection; D020798:Two-Hybrid System Techniques
D017931:DNA Primers; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D006133:Growth Substances; D018974:Insulin-Like Growth Factor Binding Protein 4; D036341:Intercellular Signaling Peptides and Proteins; D020738:Leptin; D054834:Lipocalins; D008164:Luminescent Proteins; D000077153:Progranulins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D006593:Hexokinase; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase