DOCSLIB.ORG

ON the CONTRIBUTION of BIOPHYSICAL CHARACTERIZATION to the SUCCESS in STRUCTURAL BIOLOGY Frank H

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
ON the CONTRIBUTION of BIOPHYSICAL CHARACTERIZATION to the SUCCESS in STRUCTURAL BIOLOGY Frank H ON THE CONTRIBUTION OF BIOPHYSICAL CHARACTERIZATION TO THE SUCCESS IN STRUCTURAL BIOLOGY Frank H. Niesen Structural Genomics Consortium Oxford University, Nuffield Department of Clinical Medicine Oxford, United Kingdom Madison, Dec 23, 2008 SGC Toronto SGC Oxford SGC Stockholm TOPICS Personal Profile Contributions of Biophysics to Structural Biology Projects Basics about the SGC Biophysics lab facilities Science PERSONAL PROFILE MSc in Biology (Ruhr Univ. Bochum) Yeast Cell Biology, Molecular Biology, Genetics, Plant Physiology, Biochemistry PhD in Biophysics (Humboldt-Univ. Berlin) Protein Purification (from bovine retinae), Biophysical Characterization, Enzyme Kinetics GPCR biology, G protein activation Postdoc at Protein Structure Factory (Berlin) Setup of Biophysics division, database design Protein sample optimization for crystallization Team Leader at Structural Genomics Consortium (SGC) Oxford Protein sample optimization for crystallization Functional characterization, ligand detection & characterization Molecular Biophysics, now: Chemical Biology CONTRIBUTIONS OF BIOPHYSICS TO SG Assistance in optimization of protein samples Solvent and additive screening Detection of stabilizing ligands Functional characterization for publication of structures: Quaternary structure in solution Enzymatic activity, substrate screening Characterization of protein-ligand interactions (KD, IC50, mode of binding/inhibition) Chemical Biology Structure-Activity Relationships Development of ligands into probes QUALITY MANAGEMENT IN STRUCTURAL BIOLOGY Routine characterization of properties revealed common monitors of protein quality: Identification of vibrational modes Discovery of a standard in FT-IR that are common to enthalpy of unfolding (partially) unfolded proteins J. Struct. Biol. 162:451-459 (2008) QUALITY MANAGEMENT IN STRUCTURAL BIOLOGY Concentration test combining dynamic light scattering and precipitation test on step-wise concentrated samples: Preventing loss of samples due to over- concentration Better prospect for successful crystalli- zation Study on 213 samples (98 human proteins): Type of distribution: monodisperse bimodal multimodal J. Struct. Biol. 162:451-459 (2008) Charity, i.e. not-for- THE SGC profit organization http://www.sgc.ox.ac.uk Aim: to keep basic information about protein structures in the public domain Public funding for sites in Canada, Sweden and UK Additional funding from pharma (currently: GSK, Merck, Novartis) Focus on human proteins and human disease-causing Apicomplexa Current output at Oxford site: 5 unique structures per month LAB CAPACITIES READY FOR THE JOB Liquid handling robot, for volumes < 100 nl Automated plate reader with injectors to start reactions: fluo, FP, Abs (spectra), Lumi RT-PCR machines (4) used as thermal-scanning fluorescence readers LAB CAPACITIES READY FOR THE JOB Compound Storage System Analytical Ultracentrifuge Isothermal Titration Calorimeter Dynamic Light Scattering Nanodrop Spectrometer CD spectrometer SCIENTIFIC INTERESTS Contribute to optimum in productivity and quality Sturdy methods applicable to medium-to-high throughput: Differential Scanning Fluorimetry Scripting (e.g. Excel) that enables fast and error-free analysis of large datasets Drive and organize large-scale cooperative operations: epigenetics compound screening Help discover and analyze new functional mechanisms that are common to a family or group of proteins Methods of activation: protein kinase dimerization Methods of inhibition: JmjC proteins, Tudor domains, tyrosine phosphatases Determinants of activity: Medium-chain dehydrogenase profiling Engage in high-profile single-protein projects involving mechanistic discoveries Analysis of determinants for oligomerisation: dystrophia myotonica protein kinase, discovery of additional helix in p63/p73 sequences Other single-protein projects, advising junior-level scientists in technique, strategy or interpretations DIFFERENTIAL SCANNING FLUORIMETRY Often called thermofluor; generic thermal-scanning method for the detection of stabilizing conditions, e.g. ligand binding SGC has pioneered HTS using the method, in particular with respect to the analysis (ftp://ftp.sgc.ox.ac.uk/pub/biophysics) Screening of Casein kinase 3 identified inhibitors that promote crystallization PNAS 103:15835-15840 (2006) Nature Protoc. 2(9):2212 (2007) NOVEL MECHANISM OF PROTEIN KINASE AUTO- ACTIVATION Many kinases auto-activate by inter-protein phosphorylation However, for some of them the phosphorylated region does not resemble a consensus sequence The solution: kinase domains form dimers such that the phosphorylated region rotates and points into the active site of the other protomer Dimers remain after phosphorylation, are dissociated by substrate peptide: regulation of substrate specificity possible SLK: wt dimer vs. mutant EMBO J. 27(4):704-714 (2008) ADDITIONAL HELIX IN SEQUENCES OF p63/p73 FACILITATES DIFFERENTIAL OLIGOMERIZATION AncestrAl members oF the p53 fAmIly, wIth dIstInct functIons: p63, essential for maintaining epthelIAl stem cells and protecting genomic stability; p73, involved in neuronal development And tumor suppression NMR: additional helix is only found in p63 and p73 Helix decreases dissociation constant for tetramer SLK: Helix-truncated oligomerization wt dimer domain (OD construct, red) vs. mutant is mainly dimeric (at 50 M) Submitted: Countadin et al NOVEL INHIBITOR FOR CARBONYL REDUCTASE 1 Novel method of organic synthesis called proprionate scanning: exchange of acetate by proprionate building block in certain step of synthesis of benzolactones (have vast spectrum of biological activities) First such probe, analogue of zearalenone, turned out to be very ineffective against panel of protein kinases and Hsp90 However, the Ki for carbonyl reductase 1 (major contributor to quinone reduction, creates doxrubicinol from drug targeting heart disease) lies in the nanomolar range Zearalenone ZAE analogue Bioorg. Med. Chem. , in press ACKNOWLEDGEMENTS Prof. Aled Edwards Kinase A loop dimers Helix of p63/p73 Peter Rellos, Ashley Pike (SGC) Volker Doetsch, Horng Der SGC Oxford Laurence Pearl, Anthony Oliver Ou, Daniel Countadin (Univ. Udo Oppermann (Cancer Res. UK) Frankfurt) Stefan Knapp Benjamin Turk (Yale University Differential Scanning Tom Heightman Medicine) Oleg Fedorov Fluorimetry Oliver King CBR1 Zearalenone-analogue Masoud Vedadi, Patrick Chitra Bhatia inhibitor Finnerty jr. Ron Yeung (SGC) Tabot Besong Ewa Pilka (SGC) Epigenetics Michael Sundstrøm Martin Maier, Tobias Stan Ng, Tabot Besong (SGC) Chas Bountra Zimmermann (Univ. Tübingen) Ryan Trump, Tim Willson, Ian Protein Structure Factory Characterization of CBR3 vs. Baldwin (GSK) Klaus Peter Hofmann CBR1 Anton Simeonov (NCGS) Udo Heinemann Eduard Maser, Yasser El- Chris Schofield, Nathan Rose Anja Koch, Ulf Lenski Hawari (Univ. Kiel) (Univ. Oxford) Ulrich Harttig FUNDING PARTNERS Canadian Institutes for Health Research, Canadian Foundation for Innovation, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Knut and Alice Wallenberg Foundation, Merck & Co., Inc., Novartis Research Foundation, Ontario Innovation Trust, Ontario Ministry for Research and Innovation, Swedish Agency for Innovation Systems, Swedish Foundation for Strategic Research, and Wellcome Trust. www.thesgc.org CONTACT: [email protected] +44 1865 617578 ABOUT THE SGC The SGC is an international public-private partnership (UK charity number 1097737) that aims to determine three dimensional structures of medically important human proteins, including integral membrane proteins, and proteins from human parasites. All structures, results and related reagents created by the SGC are made freely available via public databases in a prompt manner without IP constraints. www.thesgc.org.
Load more

Recommended publications
  • ANNUAL REVIEW 1 October 2005–30 September
    WELLCOME TRUST ANNUAL REVIEW 1 October 2005–30 September 2006 ANNUAL REVIEW 2006 The Wellcome Trust is the largest charity in the UK and the second largest medical research charity in the world. It funds innovative biomedical research, in the UK and internationally, spending around £500 million each year to support the brightest scientists with the best ideas. The Wellcome Trust supports public debate about biomedical research and its impact on health and wellbeing. www.wellcome.ac.uk THE WELLCOME TRUST The Wellcome Trust is the largest charity in the UK and the second largest medical research charity in the world. 123 CONTENTS BOARD OF GOVERNORS 2 Director’s statement William Castell 4 Advancing knowledge Chairman 16 Using knowledge Martin Bobrow Deputy Chairman 24 Engaging society Adrian Bird 30 Developing people Leszek Borysiewicz 36 Facilitating research Patricia Hodgson 40 Developing our organisation Richard Hynes 41 Wellcome Trust 2005/06 Ronald Plasterk 42 Financial summary 2005/06 Alastair Ross Goobey 44 Funding developments 2005/06 Peter Smith 46 Streams funding 2005/06 Jean Thomas 48 Technology Transfer Edward Walker-Arnott 49 Wellcome Trust Genome Campus As at January 2007 50 Public Engagement 51 Library and information resources 52 Advisory committees Images 1 Surface of the gut. 3 Zebrafish. 5 Cells in a developing This Annual Review covers the 2 Young children in 4 A scene from Y fruit fly. Wellcome Trust’s financial year, from Kenya. Touring’s Every Breath. 6 Data management at the Sanger Institute. 1 October 2005 to 30 September 2006. CONTENTS 1 45 6 EXECUTIVE BOARD MAKING A DIFFERENCE Developing people: To foster a Mark Walport The Wellcome Trust’s mission is research community and individual Director to foster and promote research with researchers who can contribute to the advancement and use of knowledge Ted Bianco the aim of improving human and Director of Technology Transfer animal health.
    [Show full text]
  • Holliday Junction Processing Enzymes in Eukaryotes. by Anthony John
    Holliday junction processing enzymes in eukaryotes. By Anthony John Keeley Thesis submitted to the University of London, for the degree of Doctor of Philosophy, September 1999 ProQuest Number: 10609126 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a com plete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 10609126 Published by ProQuest LLC(2017). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 ACKNOWLEDGMENTS I would like to acknowledge some of the people that without their kind help and generosity this work would have been difficult. Firstly I would like to thank members of the laboratory group, Fikret for his help, tutoring in yeast genetics, some improved methods and some long coffee breaks explaining the finer details of recombination. Judit for her help in the laboratory and always having a spare lane on a gel and Mark for his assistance with sequencing gels. I would also like to thank the other members of staff at UCL for their help and positive response to favours, Jeremy Hioms’s group, Peter Piper’s group, Laurence Pearl’s group, David Saggerson’s group, Liz Shephard’s group, Jeremy Brockes’s group and Peter Shepherd’s group.
    [Show full text]
  • Crystallography in the News Product Spotlight
    - view this in your browser - Protein Crystallography Newsletter Volume 5, No. 9, September 2013 Crystallography in the news In this issue: September 4, 2013. Computer-designed proteins that can recognize and interact with small biological molecules are now a reality. Scientists have succeeded in creating a Crystallography in the news protein molecule that can be programmed to unite with three different steroids. Crystallographers in the news September 5, 2013. While on sabbatical at the Weizmann Institute of Science in 1993, Product spotlight: Minstrel's asymmetric lighting Paul H. Axelsen—currently a professor at the University of Pennsylvania's Perelman School Lab spotlight: Pearl lab of Medicine—helped figure out how a key enzyme plays a role in communication between Useful links for crystallography certain kinds of nerve cells—the very process that sarin gas interferes with so Webinar: crystallization catastrophically. Survey of the month September 11, 2013. By taking advantage of the fact that our body proteins and robot Science video of the month arms both move in a similar way, the department of mechanical engineering of the ECM delegates raise £1376 for Cancer Research UK UPV/EHU-University of the Basque Country has developed a program to simulate protein movements. Monthly crystallographic papers Book review September 13, 2013. The Royal Swedish Academy of Sciences has awarded the Gregori Aminoff Prize in Crystallography 2014 to Yigong Shi from Tsinghua Univ. in Beijing, China for his "groundbreaking crystallographic studies of proteins and protein complexes that Crystallographers in the News regulate programmed cell death." Max Perutz Prize September 16, 2013. Stopping short of a merger, the nonprofit Hauptman-Woodward The European Crystallographic Medical Research Institute is negotiating with the University at Buffalo School of Medicine Association has awarded the seventh & Biomedical Sciences to change the way the organization and its scientists are Max Perutz Prize to Prof.
    [Show full text]
  • BRCT Domains of the DNA Damage Checkpoint Proteins
    RESEARCH COMMUNICATION BRCT domains of the DNA damage checkpoint proteins TOPBP1/Rad4 display distinct specificities for phosphopeptide ligands Matthew Day1, Mathieu Rappas1†, Katie Ptasinska2, Dominik Boos3, Antony W Oliver1*, Laurence H Pearl1* 1Cancer Research UK DNA Repair Enzymes Group, Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, United Kingdom; 2Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, United Kingdom; 3Fakulta¨ t fu¨ r Biologie, Universita¨ t Duisburg-Essen, Germany, United Kingdom Abstract TOPBP1 and its fission yeast homologue Rad4, are critical players in a range of DNA replication, repair and damage signalling processes. They are composed of multiple BRCT domains, some of which bind phosphorylated motifs in other proteins. They thus act as multi-point adaptors bringing proteins together into functional combinations, dependent on post-translational modifications downstream of cell cycle and DNA damage signals. We have now structurally and/or biochemically characterised a sufficient number of high-affinity complexes for the conserved N-terminal region of TOPBP1 and Rad4 with diverse phospho-ligands, including human RAD9 and Treslin, and Schizosaccharomyces pombe Crb2 and Sld3, to define the determinants of BRCT domain specificity. We use this to identify and characterise previously unknown phosphorylation- *For correspondence: dependent TOPBP1/Rad4-binding motifs in human RHNO1 and the fission yeast homologue of [email protected] MDC1, Mdb1. These results provide important insights into how multiple BRCT domains within (AWO); TOPBP1/Rad4 achieve selective and combinatorial binding of their multiple partner proteins. [email protected] (LHP) Editorial note: This article has been through an editorial process in which the authors decide how to respond to the issues raised during peer review.
    [Show full text]
  • Structure of an Archaeal PCNA1–PCNA2–FEN1 Complex: Elucidating PCNA Subunit and Client Enzyme Specificity Andrew S
    Published online 31 August 2006 Nucleic Acids Research, 2006, Vol. 34, No. 16 4515–4526 doi:10.1093/nar/gkl623 Structure of an archaeal PCNA1–PCNA2–FEN1 complex: elucidating PCNA subunit and client enzyme specificity Andrew S. Dore´*, Mairi L. Kilkenny, Sarah A. Jones, Antony W. Oliver, S. Mark Roe, Stephen D. Bell1 and Laurence H. Pearl* CR-UK DNA Repair Enzymes Group, Section of Structural Biology, The Institute of Cancer Research, 237 Fulham Road, Chelsea, London, SW3 6JB, UK and 1MRC Cancer Cell Unit, Hutchison MRC Research Centre, Hills Road, Cambridge, CB2 2XZ, UK Received May 2, 2006; Revised and Accepted August 8, 2006 ABSTRACT sliding clamp and vastly increasing the processivity of attached enzymes (2). The archaeal/eukaryotic proliferating cell nuclear In addition to replicative DNA polymerases, PCNA and antigen (PCNA) toroidal clamp interacts with a host its homologues have been shown to interact with a broad of DNA modifying enzymes, providing a stable range of DNA modifying enzymes implicated in such DNA anchorage and enhancing their respective proces- metabolic processes as DNA replication, Okazaki fragment sivities. Given the broad range of enzymes with maturation, DNA methylation, nucleotide excision repair which PCNA has been shown to interact, relatively (NER), mismatch repair (MMR) and long-patch base excision little is known about the mode of assembly of func- repair (LP-BER). In general, the principal interaction is tionally meaningful combinations of enzymes on the mediated by recognition of a PCNA-interacting protein PCNA clamp. We have determined the X-ray crystal motif (PIP-box) of the ‘client’ enzyme, which binds the inter- structure of the Sulfolobus solfataricus PCNA1– domain connector loop (IDCL) of a PCNA subunit (3).
    [Show full text]
  • Protein Structural Analysis: Α-Helices and Their Interactions
    Protein Structural Analysis: a-helices and their interactions by Frances Mary Genevieve Pearl Biomolecular and Structural Modelling Unit, Department of Biochemistry and Molecular Biology, University College London, Gower Street, London. WCIE 6BT This thesis is submitted in fulfilment of the requirements for the degree of Doctor of Philosophy from the University of London March 1998 ProQuest Number: U643156 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest. ProQuest U643156 Published by ProQuest LLC(2016). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code. Microform Edition © ProQuest LLC. ProQuest LLC 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106-1346 Abstract Abst r a c t Helices are fundamental building blocks of many protein structures, and interactions between them play a crucial role in stabilising a protein’s fold. This work updates and enhances the current knowledge of the nature of these interactions using the 3-dimensional coordinates of protein structures deposited in the Protein Data Bank. Having defined rigorous criteria for identifying regular a-helices, the geometric and physical properties of helix-helix interactions were studied. In contrast with previous studies, a systematic analysis of a large dataset of helix-helix interactions was undertaken.
    [Show full text]
  • Chapter 7 Replication Checkpoint Dependent Rad4 Interactions
    A University of Sussex DPhil thesis Available online via Sussex Research Online: http://sro.sussex.ac.uk/ This thesis is protected by copyright which belongs to the author. This thesis cannot be reproduced or quoted extensively from without first obtaining permission in writing from the Author The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the Author When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given Please visit Sussex Research Online for more information and further details Protein-Protein Interactions Underlying Damage Checkpoint Activation in S. pombe Christopher Wardlaw Submitted for the Degree of Doctor of Philosophy University of Sussex September 2013 II Declaration I hereby declare that this thesis has not been and will not be, submitted in whole or in part to another University for the award of any other degree. Signature:……………………………………… III Acknowledgements I would like to thank Tony Carr for the opportunity to undertake my PhD in his laboratory, and for his great supervision, help and advice throughout the process. I would also like to thank Jo Murray for her helpful input, ideas and discussion, and thank my second supervisor Alessandro Bianchi. Many thanks to all of the Carr and Murray lab members, past and present, for their help and input over the past four years, and for creating such a wonderful working environment. Special thanks to Valerie Garcia for her supervision and support, and Adam Watson for his never ending technical help and patience.
    [Show full text]
  • Green Biotechnology: Kill Or Cure?
    issue 11 winter 2008|2009 promoting excellence in the molecular life sciences in europe Dear Reader, Green biotechnology: kill or cure? Recognising excellence is core to EMBO – the member- ship has been doing just that since nomination of the fi rst 200 EMBO Members in the 1960’s. This year again we welcome newly elected members to EMBO (see page 4). And we congratulate Luc Montagnier, Roger Tsien and Harald zur Hausen – EMBO Members awarded The Nobel Prize this year. The discipli- nary breadth of molecular life sciences is much broader today than it was some 40 years ago. For this reason, a modifi ed member election procedure was adopted – see page 5. You may have noticed some changes to format in this issue of EMBOencounters: fi rst, you are hearing from me as Deputy Director – a role I share with EMBO Fellowships Programme Manager Jan Taplick. Secondly, we plan a lead story for each issue to highlight © www.goldenrice.org topics relevant to EMBO activities. Golden Rice could help prevent vitamin A defi ciency in the developing world. This issue’s lead story investigates today’s Soaring grain prices, high energy costs and increasingly louder riots on the streets of perceptions of the green revolution – the focus famine-stricken countries such as Haiti or Somalia are forcing politicians and the public of the next EMBO/EMBL Science & Society to reconsider their opposition to modern agriculture and crops created through breed- Conference. EMBO Science & Society aims to ing techniques that employ methods of molecular genetics. Will the fi erce opposition of create dialogue between policy makers and western countries to so-called genetically modifi ed (GM) crops eventually give way to the public and complements our numerous the acceptance that they might help tackle the global food crisis and even prevent some activities that share knowledge addressing the diseases? challenges of our changing world.
    [Show full text]
  • ATP-Competitive Inhibitors Block Protein Kinase Recruitment to the Hsp90-Cdc37 System
    Europe PMC Funders Group Author Manuscript Nat Chem Biol. Author manuscript; available in PMC 2017 November 20. Published in final edited form as: Nat Chem Biol. 2013 May ; 9(5): 307–312. doi:10.1038/nchembio.1212. Europe PMC Funders Author Manuscripts ATP-competitive inhibitors block protein kinase recruitment to the Hsp90-Cdc37 system Sigrun Polier1,*, Rahul S. Samant2, Paul A. Clarke2, Paul Workman2,*, Chrisostomos Prodromou1, and Laurence H. Pearl1,* 1MRC Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 9RQ, UK 2Cancer Research UK Cancer Therapeutics Unit, The Institute of Cancer Research, London SM2 5NG, UK Abstract Protein kinase clients are recruited to the Hsp90 molecular chaperone system via Cdc37, which simultaneously binds Hsp90 and kinases and regulates the Hsp90 chaperone cycle. Pharmacological inhibition of Hsp90 in vivo results in degradation of kinase clients, with a therapeutic effect in dependent tumours. We show here that Cdc37 directly antagonises ATP binding to client kinases, suggesting a role for the Hsp90-Cdc37 complex in controlling kinase activity. Unexpectedly, we find that Cdc37 binding to protein kinases is itself antagonised by ATP- competitive kinase inhibitors including vemurafenib and lapatinib. In cancer cells these inhibitors deprive oncogenic kinases such as BRaf and ErbB2 of access to the Hsp90-Cdc37 complex, Europe PMC Funders Author Manuscripts leading to their degradation. Our results suggest that at least part of the efficacy of ATP- competitive inhibitors of Hsp90-dependent kinases clients in tumour cells may be due to targeted chaperone deprivation. Introduction Protein kinases, which function as the major regulators and transducers of signalling in eukaryotic cells, constitute the largest coherent class of client proteins of the Hsp90 molecular chaperone system 1.
    [Show full text]
  • Annual Report and Financial Statements for the Year Ended 31 July 2019
    Annual Report and Financial Statements for the year ended 31 July 2019 The Institute of Cancer Research: Royal Cancer Hospital Company Number 00534147 2019 2 Contents Executive summary 4–9 1 Report of the Board of Trustees 11–45 Objectives and activities 12–15 Strategic report 17–37 Governance and management 39–42 Statement of the responsibilities of members 44–45 of the Board of Trustees 2 Independent Auditor’s Report 47–51 3 The Financial Statements 53–79 Consolidated and ICR statement of comprehensive income and expenditure 54 Consolidated and ICR statement of changes in reserves 55 Consolidated and ICR balance sheets 56 Consolidated statement of cashflows 57 Statement of accounting policies 58–62 Notes to the financial statements 63–79 4 ICR information 81–86 The Board of Trustees 82 Governing committees, fellows, members and associates 83–85 Legal and administrative information 86 The Institute of Cancer Research: Royal Cancer Hospital Company Number 00534147 Financial Statements for the year ended 31 July 2019 3 Executive summary Our finances In 2018/19 The Institute of Cancer Research, London, had total income of £167.4m. We received 40% of our income from research £167.4m grants, 27% in public funding as a higher of income in 2018/19 education institution, 22% from royalty income, 7% from donations and endowments, and 4% from tuition fees, investments and other income. Expenditure was £143.3m, of which 78% was spent directly on research. We spent 17% on supporting this research by creating £143.3m the best possible environment for our of operating expenditure in 2018/19 scientists, 3% on fundraising and 2% on information and other activities.
    [Show full text]
  • Specialized Interfaces of Smc5/6 Control Hinge Stability and DNA Association
    ARTICLE Received 7 Jul 2016 | Accepted 21 Nov 2016 | Published 30 Jan 2017 DOI: 10.1038/ncomms14011 OPEN Specialized interfaces of Smc5/6 control hinge stability and DNA association Aaron Alt1, Hung Q. Dang2, Owen S. Wells2, Luis M. Polo1, Matt A. Smith2, Grant A. McGregor2, Thomas Welte3, Alan R. Lehmann2, Laurence H. Pearl1, Johanne M. Murray2 & Antony W. Oliver1 The Structural Maintenance of Chromosomes (SMC) complexes: cohesin, condensin and Smc5/6 are involved in the organization of higher-order chromosome structure—which is essential for accurate chromosome duplication and segregation. Each complex is scaffolded by a specific SMC protein dimer (heterodimer in eukaryotes) held together via their hinge domains. Here we show that the Smc5/6-hinge, like those of cohesin and condensin, also forms a toroidal structure but with distinctive subunit interfaces absent from the other SMC complexes; an unusual ‘molecular latch’ and a functional ‘hub’. Defined mutations in these interfaces cause severe phenotypic effects with sensitivity to DNA-damaging agents in fission yeast and reduced viability in human cells. We show that the Smc5/6-hinge complex binds preferentially to ssDNA and that this interaction is affected by both ‘latch’ and ‘hub’ mutations, suggesting a key role for these unique features in controlling DNA association by the Smc5/6 complex. 1 Cancer Research UK DNA Repair Enzymes Group, Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, BN1 9RQ, UK. 2 Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9RQ, UK. 3 Dynamic Biosensors GmbH, Lochhamer Strasse, D-81252 Martinsreid/Planegg, Germany.
    [Show full text]
  • In Vitro Biological Characterization of a Novel, Synthetic Diaryl Pyrazole Resorcinol Class of Heat Shock Protein 90 Inhibitors
    Research Article In vitro Biological Characterization of a Novel, Synthetic Diaryl Pyrazole Resorcinol Class of Heat Shock Protein 90 Inhibitors Swee Y. Sharp,1 Kathy Boxall,1 Martin Rowlands,1 Chrisostomos Prodromou,2 S. Mark Roe,2 Alison Maloney,1 Marissa Powers,1 Paul A. Clarke,1 Gary Box,1 Sharon Sanderson,1 Lisa Patterson,1 Thomas P. Matthews,1 Kwai-Ming J. Cheung,1 Karen Ball,1 Angela Hayes,1 Florence Raynaud,1 Richard Marais,3 Laurence Pearl,2 Sue Eccles,1 Wynne Aherne,1 Edward McDonald,1 and Paul Workman1 1Haddow Laboratories, Cancer Research UK Centre for Cancer Therapeutics, The Institute of Cancer Research, Surrey, United Kingdom; 2Chester Beatty Laboratories, Section of Structural Biology; and 3Cancer Research UK Centre for Cell and Molecular Biology, The Institute of Cancer Research, London, United Kingdom Abstract Introduction The molecular chaperone heat shock protein 90 (HSP90) has The molecular chaperone heat shock protein 90 (HSP90) emerged as an exciting molecular target. Derivatives of the regulates the stability, activation, and biological function of natural product geldanamycin,such as 17-allylamino-17- numerous oncogenic client proteins, including steroid hormone demethoxy-geldanamycin (17-AAG),were the first HSP90 receptors, kinases (e.g., ERBB2, C-RAF, B-RAF, and CDK4), and ATPase inhibitors to enter clinical trial. Synthetic small- other proteins (e.g., mutant p53, hTERT, and HIF1a; ref. 1). The molecule HSP90 inhibitors have potential advantages. Here, HSP90 chaperone machine is driven by ATP binding and hydrolysis we describe the biological properties of the lead compound (2). Its activity is regulated by co-chaperones (e.g., HSP72, CDC37, of a new class of 3,4-diaryl pyrazole resorcinol HSP90 inhib- P23, CHIP, and immunophilins) that affect the balance between itor (CCT018159),which we identified by high-throughput stabilization and degradation of clients via the ubiquitin-protea- screening.
    [Show full text]