Methods Relating to Mammalian Rictor Polypeptide
AssigneeWhitehead Institute for Biomedical Research
InventorsDavid M. Sabatini, Do-Hyung Kim, Dos D. Sarbassov
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Abstract
The invention describes isolated mTOR-associated proteins (“mTOR-APs”) as well as isolated variants and fragments thereof and the isolated nucleic acids encoding them. The invention also describes vectors and host cells containing nucleic acid encoding an mTOR-AP polypeptide and methods for producing an mTOR-AP polypeptide. Also described are methods for screening for compounds which modulate mTOR-AP activity and methods for treating or preventing a disorder that is responsive to mTOR-AP modulation.
Claims12
A method for identifying a compound which modulates the activity of a rictor polypeptide, comprising:
- (a) contacting an isolated cell or cell lysate with a test agent, wherein the cell or cell lysate comprises:
- (i) a polypeptide comprising a rictor amino acid sequence that is at least 97% identical to SEQ ID NO:
- 3, wherein the rictor amino acid sequence can form a complex with human mTOR and can reduce complex formation between raptor and human mTOR;
- and
- (ii) human mTOR or a fragment thereof, wherein the human mTOR protein, or fragment thereof, forms a complex with the rictor amino acid sequence;
- and
- (b) measuring:
- (i) the amount of the complex comprising the rictor amino acid sequence and human mTOR in the cell or cell lysate;
- or
- (ii) the amount of mTOR kinase activity of the complex in the cell or cell lysate, wherein mTOR kinase activity is measured by analysis of phosphorylation of a human mTOR substrate;
- and
- (c) comparing the measurement of step (b) with a measurement obtained for a control sample without the test agent;
- wherein a compound which modulates the activity of the rictor polypeptide is identified if the measurement of the amount of the complex or the amount of mTOR kinase activity in step (b) varies from a measurement obtained for the control sample without the test agent.
Dependencies are inferred from the claim text.