Germline Mutations in the CDKN2B Tumor Suppressor Gene Predispose to Renal Cell Carcinoma.
Cancer Discov. 2015 Jul;5(7):723-9
Authors: Jafri M, Wake NC, Ascher DB, Pires DE, Gentle D, Morris MR, Rattenberry E, Simpson MA, Trembath RC, Weber A, Woodward ER, Donaldson A, Blundell TL, Latif F, Maher ER
UNLABELLED: Familial renal cell carcinoma (RCC) is genetically heterogeneous and may be caused by mutations in multiple genes, including VHL, MET, SDHB, FH, FLCN, PTEN, and BAP1. However, most individuals with inherited RCC do not have a detectable germline mutation. To identify novel inherited RCC genes, we undertook exome resequencing studies in a familial RCC kindred and identified a CDKN2B nonsense mutation that segregated with familial RCC status. Targeted resequencing of CDKN2B in individuals (n = 82) with features of inherited RCC then revealed three candidate CDKN2B missense mutations (p.Pro40Thr, p.Ala23Glu, and p.Asp86Asn). In silico analysis of the three-dimensional structures indicated that each missense substitution was likely pathogenic through reduced stability of the mutant or reduced affinity for cyclin-dependent kinases 4 and 6, and in vitro studies demonstrated that each of the mutations impaired CDKN2B-induced suppression of proliferation in an RCC cell line. These findings identify germline CDKN2B mutations as a novel cause of familial RCC.
SIGNIFICANCE: Germline loss-of-function CDKN2B mutations were identified in a subset of patients with features of inherited RCC. Detection of germline CDKN2B mutations will have an impact on familial cancer screening and might prove to influence the management of disseminated disease.
PMID: 25873077 [PubMed - indexed for MEDLINE]
Identifying Interactions that Determine Fragment Binding at Protein Hotspots.
J Med Chem. 2016 Apr 4;
Authors: Radoux CJ, Olsson TS, Pitt WR, Groom CR, Blundell TL
Locating a ligand binding site is an important first step in structure-guided drug discovery, but current methods do little to suggest which interactions within a pocket are the most important for binding. Here we illustrate a method that samples atomic hotspots with simple molecular probes to produce fragment hotspot maps. These maps specifically highlight fragment binding sites and their corresponding pharmacophores. For ligand-bound structures, they provide an intuitive visual guide within the binding site, directing medicinal chemists where to grow the molecule and alerting them to sub-optimal interactions within the original hit. The fragment hotspot map calculation is validated using experimental binding positions of 21 fragments and subsequent lead molecules. The ligands are found in high scoring areas of the fragment hotspot maps, with fragment atoms having a median percentage rank of 97%. Protein kinase B and pantothenate synthetase are examined in detail. In each case the fragment hotspot maps are able to rationalise a Free-Wilson analysis of SAR data from a fragment-based drug design project.
PMID: 27043011 [PubMed - as supplied by publisher]
Structure Activity Relationship of the Peptide Binding Motif Mediating the RAD51:BRCA2 Protein-Protein Interaction.
FEBS Lett. 2016 Mar 19;
Authors: Scott DE, Marsh M, Blundell TL, Abell C, Hyvönen M
RAD51 is a recombinase involved in the homologous recombination of double strand breaks in DNA. RAD51 forms oligomers by binding to another molecule of RAD51 via an "FxxA" motif, and the same recognition sequence is similarly utilized to bind BRCA2. We have tabulated the effects of mutation of this sequence, across a variety of experimental methods and from relevant mutations observed in the clinic. We use mutants of a tetrapeptide sequence to probe the binding interaction, using both isothermal titration calorimetry and X-ray crystallography. Where possible, comparison between our tetrapeptide mutational study and the previously reported mutations is made, discrepancies are discussed, and the importance of secondary structure in interpreting alanine-scanning and mutational data of this nature is considered. This article is protected by copyright. All rights reserved.
PMID: 26992456 [PubMed - as supplied by publisher]