Structure of a complex of retinoblastoma protein bound to e2f, and uses thereof
OwnerSteven Gamblininventor
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Abstract
The present invention provides the crystal structure of pRb/E2F(409-426) as well as uses of the structure in identifying agents which modulate the binding between pRb and E2F and/or a pRb/E2F(409-426) complex, and thus are useful as pharmaceutical agents in the prevention or treatment of proliferative diseases.
Claims47
A crystal comprising a pRb/E2F(409-426) complex, wherein the crystal structure is characterised by the atomic co-ordinates of Annex 1.
A method of identifying an agent which modulates the interaction between pRb and E2F(409-426), comprising:
- a) combining pRb, E2F(409-426) and an agent, under conditions in which pRb and E2F(409-426) form a complex;
- b) growing a crystal structure of any pRb/ E2F(409-426) complex;
- and
- c) analyzing the crystal to determine whether the agent modulates the interaction between pRb and E2F(409-426).
A method of identifying an agent that modulates a pRb/E2F(409-426) complex, comprising selecting an agent using the three-dimensional atomic coordinates of Annex 1.
A method of identifying an agent that modulates a pRb/E2F(409-426) complex, comprising:
- a) contacting an agent with pRb and E2F(409-426) under conditions in which pRb and E2F(409-426) can form a complex;
- and
- b) measuring the binding affinity of pRb to E2F(409-426) in the presence of the agent and comparing the binding affinity to that of pRb to E2F(409-426) when in the absence of the agent, wherein an agent modulates a pRb/E2F(409-426) complex when there is a change in the binding affinity of pRb to E2F(409-426) when in the presence of the agent.
A method of identifying an agent that modulates a pRb/E2F(409-426) complex, comprising:
- a) contacting a fluorescently tagged E2F(409-426) peptide (E2F-fluoropeptide) with pRb to allow pRb/E2F-fluoropeptide complex formation;
- b) detecting the fluorescence polarization;
- c) adding an agent;
- and
- d) detecting the fluorescence polarization in the presence of the agent.
A method of identifying an agent that modulates a pRb/E2F(409-426) complex, comprising;
- a) contacting a fluorescently tagged E2F(409-426) peptide (E2F-fluoropeptide) with pRb to allow pRb/E2F-fluoropeptide complex formation;
- b) detecting the fluorescence polarization;
- c) contacting an agent with pRb and E2F(409-426) peptide (E2F-fluoropeptide) under conditions in which pRb and E2F-fluoropeptide can form a complex;
- d) detecting the fluorescence polarization;
- and
- e) comparing the fluorescence polarization detected in b) and d).
(canceled)
Computer readable media comprising a data storage material encoded with computer readable data, wherein said computer readable data comprises a set of atomic co-ordinates of the pRb/E2F(409-426) crystal structure of Annex I recorded thereon.
Dependencies are inferred from the claim text.