Nucleic acids encoding a mammalian raptor polypeptide and uses therefor
AssigneeWhitehead Institute for Biomedical Research
InventorsDavid M. Sabatini, Do-Hyung Kim, Dos Sarbassov
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Abstract
The present invention relates to isolated raptor nucleic acid molecules of mammalian origin (e.g., human) and complements, portions and variants thereof. Another aspect of the invention are isolated raptor polypeptides of mammalian origin and portions thereof, and antibodies or antigen binding fragments thereof that specifically bind a raptor polypeptide. The present invention also relates to constructs and host cells comprising the nucleic acid molecules described herein. In addition, the present invention relates to uses of the nucleic acid and polypeptide molecules provided herein.
Claims53
An isolated nucleic acid molecule comprising SEQ ID NO:
- 1.
An isolated nucleic acid molecule which is the complement of SEQ ID NO:
- 1.
An isolated nucleic acid molecule that encodes an amino acid sequence comprising SEQ ID NO:
- 2.
An isolated nucleic acid molecule comprising a sequence that hybridizes under highly stringent conditions to SEQ ID NO:
- 1 or a complement of SEQ ID NO:
- 1.
An isolated nucleic acid molecule comprising a sequence that hybridizes under highly stringent conditions to a complement of SEQ ID NO:
- 1 and encodes a mammalian raptor protein.
An isolated polypeptide encoded by a nucleic acid comprising SEQ ID NO:
- 1.
An isolated polypeptide having an amino acid sequence comprising SEQ ID NO:
- 2.
An expression construct comprising SEQ ID NO:
- 1.
An antibody or antigen binding fragment thereof that specifically binds to a mammalian raptor polypeptide, wherein the mammalian raptor polypeptide is encoded by an isolated nucleic acid that encodes SEQ ID NO:
- 2.
A method of identifying a nucleic acid that encodes a mammalian raptor polypeptide in a sample comprising:
- a) contacting the sample with a nucleotide sequence comprising SEQ ID NO:
- 1 under conditions in which hybridization occurs between SEQ ID NO:
- 1 and nucleic acid in the sample using high stringency conditions;
- b) identifying the nucleic acid of a) which hybridizes to SEQ ID NO:
- 1 under high stringency conditions,
- thereby identifying a nucleic acid that encodes a mammalian raptor polypeptide in a sample.
A method of identifying a mammalian raptor polypeptide in a sample comprising:
- a) contacting the sample with an antibody or antigen binding fragment thereof that specifically binds to a mammalian raptor polypeptide wherein the mammalian raptor polypeptide is encoded by an isolated nucleic acid that encodes SEQ ID NO:
- 2;
- and
- b) identifying the polypeptide which specifically binds to the antibody, thereby identifying a mammalian raptor polypeptide in a sample.
A method of identifying an agent that alters interaction of a mammalian raptor protein with mammalian target of rapamycin (mTOR) protein comprising:
- a) contacting a raptor protein having an amino acid sequence comprising SEQ ID NO:
- 2 with mTOR protein under conditions in which the raptor protein interacts with the mTOR protein, with an agent to be assessed;
- b) assessing the extent to which raptor interacts with mTOR in the presence of the agent to be assessed,
- wherein if the extent to which raptor interacts with mTOR is altered in the presence of the agent compared to the extent to which raptor interacts with mTOR in the absence of the agent, then the agent alters interaction of a mammalian raptor protein with mTOR protein.
A method of identifying an agent that alters interaction of a mammalian raptor protein with mammalian target of rapamycin (mTOR) protein comprising:
- a) contacting a cell which comprises nucleic acid that encodes a raptor protein having an amino acid sequence comprising SEQ ID NO:
- 2 wherein the raptor protein, when expressed, interacts with mTOR protein in the cell, with an agent to be assessed;
- b) assessing growth rate or size of the cell,
- wherein an altered growth rate or size of the cell compared to the growth rate or size of a control cell indicates that the agent alters interaction of a mammalian raptor protein with mTOR protein.
A method of identifying an agent that inhibits an interaction of a mammalian raptor protein with mammalian target of rapamycin (mTOR) protein comprising:
- a) contacting a cell which comprises nucleic acid that encodes a raptor protein having an amino acid sequence comprising SEQ ID NO:
- 2 wherein the raptor protein, when expressed, interacts with mTOR protein in the cell, with an agent to be assessed;
- b) assessing growth rate or size of the cell,
- wherein a decrease in growth rate or size of the cell compared to growth rate or size of a control cell indicates that the agent inhibits interaction of a mammalian raptor protein with mTOR protein.
A method of identifying an agent that enhances an interaction of a mammalian raptor protein with mammalian target of rapamycin (mTOR) protein comprising:
- a) contacting a cell which comprises nucleic acid that encodes a raptor protein having an amino acid sequence comprising SEQ ID NO:
- 2 wherein the raptor protein, when expressed, interacts with mTOR protein in the cell, with an agent to be assessed;
- b) assessing growth rate or size of the cell,
- wherein an increase in growth rate or size of the cell compared to growth rate and/or size of a control cell indicates that the agent enhances interaction of a mammalian raptor protein with mTOR protein.
A method of altering growth rate or size of a cell comprising introducing into the cell an agent that alters interaction of mammalian raptor protein with mTOR.
A method of enhancing growth rate or size of a cell comprising introducing into the cell an agent that inhibits interaction of mammalian raptor protein with mammalian TOR protein.
A method of inhibiting growth rate or size of a cell comprising introducing into the cell an agent that enhances interaction of mammalian raptor protein with mammalian TOR protein.
A method of altering kinase activity of mTOR protein in a cell comprising introducing into the cell an agent that alters interaction of a mammalian raptor protein with the mTOR.
A method of enhancing kinase activity of mTOR protein in a cell comprising introducing into the cell an agent that inhibits interaction of a mammalian raptor protein with the mTOR.
A method of inhibiting kinase activity of mTOR protein in a cell comprising introducing into the cell an agent that enhances interaction of a mammalian raptor protein with the mTOR.
A method of treating cancer in an individual comprising administering to the individual an agent that enhances interaction of raptor protein with mTOR protein.
A method of identifying proteins that associate with mTOR within a cell comprising:
- a) contacting a lysate of the cell with a reversible crosslinker thereby forming a crosslink between proteins that associate with mTOR within the cell;
- b) separating proteins that are associated by the crosslink from the lysate;
- thereby identifying proteins that associate with mTOR within a cell.
A method of identifying proteins that associate with mTOR within a cell, comprising:
- a) preparing a lysate of the cell with a buffer comprising a detergent other than Triton, thereby preserving an association between the proteins;
- b) separating proteins that are associated from the lysate, thereby identifying proteins that associate with mTOR within a cell.
Dependencies are inferred from the claim text.