Polypeptide methods and means
OwnerAshok Venkitaraman, Luca Pellegrini +3inventors
InventorsAshok Venkitaraman, Luca Pellegrini, Tom Blundell, David Yu, Debbie Bates
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Abstract
The structure of a RAD51-BRC repeat sequence complex structure is provided. The structure can be used in modelling the interaction of molecular structures such as potential pharmaceutical compounds. Mutant RAD51 and BRCA2 polypeptides and RAD51-BRC repeat sequence chimaera proteins and are also provided. The mutants may be used in assays for finding compounds which interact with or form part of a RAD51 pathway, and the chimaeras can be used to form crystals which may be analysed by X-ray crystallography.
Claims18
1-19. (canceled)
A crystal of a RAD51-BRC repeat sequence complex.
A crystal according to claim 1 having the orthorhombic space group P212121, and unit cell dimensions a=57.30 ű5%, b=59.14 ű5%, c=77.20 ű5%.
A RAD51-BRC repeat sequence chimaera protein in which the RAD51 is covalently joined to the BRC repeat sequence.
A RAD51 paralogue-BRC repeat sequence chimaera protein in which the RAD51 paralogue is covalently joined to the BRC repeat sequence.
A mutant RAD51 which has been modified to reduce or eliminate the tendency of RAD51 to spontaneously aggregate into high molecular weight complexes.
A mutant RAD51 which has been modified by substitution, deletion and/or addition of at least one amino acid in the 85-GFTTATE-91 sequence of human RAD51, or the corresponding sequence in other forms of RAD51.
A method of homology modelling comprising the steps of:
- (a) aligning a representation of an amino acid sequence of a target protein of unknown three-dimensional structure with the amino acid sequence of the RAD51 or the BRC repeat sequence of Table 1 to match homologous regions of the amino acid sequences;
- (b) modelling the structure of the matched homologous regions of said target protein of unknown structure on the corresponding regions of the RAD51 or BRC repeat sequence structure as defined by Table 1;
- and
- (c) determining a conformation for said target protein of unknown structure which substantially preserves the structure of said matched homologous regions.
A method for determining the structure of a protein, which method comprises;
- providing the co-ordinates of Table 1, and
- positioning the co-ordinates in the crystal unit cell of said protein so as to provide a structure for said protein.
A method for determining the structure of a compound bound to RAD51 or a BRC repeat sequence, said method comprising:
- providing a crystal of a complex in which a compound is bound to RAD51 or a BRC repeat sequence;
- and
- determining the structure of said complex by employing the data of Table 1.
A computer-based method for the analysis of the interaction of a molecular structure with RAD51 or BRC repeat sequence, which comprises:
- providing the structure of RAD51 or a BRC repeat sequence as defined by Table 1;
- providing a molecular structure to be fitted to said RAD51 or BRC repeat sequence structure;
- and
- fitting the molecular structure to the RAD51 or BRC repeat sequence structure.
A computer-based method for the analysis of the interaction of a molecular structure with RAD51 or BRC repeat sequence, which comprises:
- providing the coordinates of at least two atoms of RAD51 or a BRC repeat sequence structure as defined by Table 1;
- providing a molecular structure to be fitted to said coordinates;
- and
- fitting the structure to the said coordinates.
Dependencies are inferred from the claim text.