Gene interactions and pathways from curated databases and text-mining
Biochem Biophys Res Commun 2005, PMID: 15896331

Characterization of the cloned full-length and a truncated human target of rapamycin: activity, specificity, and enzyme inhibition as studied by a high capacity assay.

Toral-Barza, Lourdes; Zhang, Wei-Guo; Lamison, Craig; Larocque, James; Gibbons, James; Yu, Ker

The mammalian target of rapamycin (mTOR/TOR) is implicated in cancer and other human disorders and thus an important target for therapeutic intervention. To study human TOR in vitro, we have produced in large scale both the full-length TOR (289 kDa) and a truncated TOR (132 kDa) from HEK293 cells. Both enzymes demonstrated a robust and specific catalytic activity towards the physiological substrate proteins, p70 S6 ribosomal protein kinase 1 (p70S6K1) and eIF4E binding protein 1 (4EBP1), as measured by phosphor-specific antibodies in Western blotting. We developed a high capacity dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) for analysis of kinetic parameters. The Michaelis constant (Km) values of TOR for ATP and the His6-S6K substrate were shown to be 50 and 0.8 microM, respectively. Dose-response and inhibition mechanisms of several known inhibitors, the rapamycin-FKBP12 complex, wortmannin and LY294002, were also studied in DELFIA. Our data indicate that TOR exhibits kinetic features of those shared by traditional serine/threonine kinases and demonstrate the feasibility for TOR enzyme screen in searching for new inhibitors.

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Text Mining Data

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Manually curated Databases

  • IRef Biogrid Interaction: PA2G4 — MTOR (direct interaction, pull down)
  • IRef Biogrid Interaction: RPS6KB1 — MTOR (direct interaction, pull down)
In total, 2 gene pairs are associated to this article in curated databases