Identification and purification of a soluble region in the breast cancer susceptibility protein BRCA2.
The BRCA2 gene encodes a large multidomain protein of 3418 residues. Studies to elucidate the mechanisms by which BRCA2 prevents tumour formation have been largely restricted by the size of the protein. Based on secondary structure predictions we have cloned regions across the BRCA2 gene and determined the solubility of the proteins they encode. The fragment consisting of amino acids 290-456 BRCA2 was found predominantly in the soluble portion of the cell lysate and was purified by ion exchange and nickel-NTA affinity chromatography. CD spectroscopy revealed secondary structure elements consistent with a folded peptide and limited proteolysis was used to identify a potential novel domain.
Finch
David
D
Department of Medical Genetics, Centre for Molecular Medicine, University of Manchester, Stopford Building, Manchester M13 9PT, UK.
Webb
Michelle
M
D000595:Amino Acid Sequence; D024682:BRCA2 Protein / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D002942:Circular Dichroism; D003001:Cloning, Molecular; D015870:Gene Expression; D020022:Genetic Predisposition to Disease; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D017510:Protein Folding; D012995:Solubility
D024682:BRCA2 Protein; D010446:Peptide Fragments