Structural studies of phosphoinositide signalling and intracellular sorting

Lead Research Organisation: MRC Laboratory of Molecular Biology

Abstract

Mammalian cells have a complex regulation that enables them to respond to growth factors, neurotransmitters and hormones. However, they also have cell-intrinsic responses to starvation and other environmental stresses. We are determining structures and dynamics of the cellular machines that integrate these cues to regulate cell responses. Lipid membranes partition organelles within cells and they help organise these responses. An important component of the work we do concerns how cellular machines interact with these membranes and how the shape and composition of the membranes determine the activity of these molecular machines. We are using the structures and dynamics of these machines to contribute to development of pharmaceuticals that may be useful in treating cancer, inflammation, diabetes and ageing.

Technical Summary

Eukaryotic cells have evolved a number of mechanisms to respond to environmental stress. These stress responses are tempered by growth factors and other signals from surrounding cells in multicellular organisms. Our research programme involves the mechanisms whereby these cellular signals are coupled to regulation of anabolic and catabolic processes in mammalian cells. Phosphoinositide 3-kinases (PI3Ks) modify lipid membranes and the modified phospholipids produced by the enzymes serve as second messengers and sorting signals in eukaryotic cells. The PI3Ks are essential for cell response to environmental cues. Furthermore, PI3Ks are part of a larger family of kinases known as the PI3K-related kinases (PIKKs). We study the structures, dynamics and functions of PIKKs and related pathways in cellular signalling, sorting, stress response and DNA damage repair. This has included almost all of the members of the PI3K family: class I PI3Ks activated by receptor tyrosine kinase and GPCRS, the primordial class III PI3K complex present in all eukaryotes, the golgi-associated PI4K, the PI3K related protein kinases (PIKKs) mTOR, ATM and ATR. Each of these PIKKs is a regulatory nexus, and our work produces variants in which specific regulatory inputs of the large enzyme complexes are ablated without affecting other activities. These modified complexes provide tools to decipher cellular regulation in ways that would not be possible with other approaches. We use X-ray crystallography, cryo-electron microscopy (cryo-EM) and hydrogen-deuterium exchange mass spectrometry (HDX-MS) to marry structures with dynamics. In collaboration with our pharmaceutical partners, we are seeking to develop HDX-MS approaches to give better insight into conformational dynamics that could result in new therapeutic strategies. On-going efforts in the group concern the structural and dynamic basis of regulation of oncogenic PI3Ks, autophagic PI3K complexes, integration of amino acid and growth factor sensing in the PI3K pathway, regulation of ribosomes in response to amino acid starvation and DNA damage repair.

Publications

10 25 50
 
Description Chairman Scientific Advisory Board, PIQUR Therapeutics
Geographic Reach Europe 
Policy Influence Type Participation in a advisory committee
 
Description Diamond Light source peer review panel
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Member of the MRC Molecular and Cellular Medicine Board
Geographic Reach National 
Policy Influence Type Membership of a guideline committee
 
Description Scientific advisory board for max planck society, dortmund
Geographic Reach Asia 
Policy Influence Type Participation in advisory committee
 
Description AstraZeneca-LMB Collaboration on HDX-MS Fragmentation
Amount £300,000 (GBP)
Organisation AstraZeneca 
Sector Private
Country United Kingdom
Start 04/2015 
End 04/2018
 
Description AstraZeneca/LMB Blue-Skies Inniative. Structures of ATM and ATR complexes
Amount £200,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 09/2015 
End 08/2017
 
Description BBSRC 2012 Responsive Mode Grant
Amount £120,465 (GBP)
Funding ID BB/K019155/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 05/2013 
End 04/2016
 
Description Cambridge Cancer Centre Studentship (Rotislavleva)
Amount £80,000 (GBP)
Organisation Cancer Research UK Cambridge Institute 
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 09/2016
 
Description Cancer Research UK Science Committee - Programme Award
Amount £384,252 (GBP)
Funding ID C14801/A21211 
Organisation Cancer Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2016 
End 02/2021
 
Description EMBO Long Term Fellowship (Burke)
Amount £60,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Learned Society
Country European Union (EU)
Start 01/2010 
End 12/2011
 
Description EMBO Long Term Fellowship (Pobbati)
Amount £48,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Learned Society
Country European Union (EU)
Start 09/2007 
End 09/2009
 
Description EMBO Long Term Fellowship (Sachse)
Amount £45,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Learned Society
Country European Union (EU)
Start 06/2008 
End 03/2010
 
Description EMBO Long Term Fellowship (Vadas)
Amount £7,500 (GBP)
Organisation European Molecular Biology Organisation 
Sector Learned Society
Country European Union (EU)
Start 09/2010 
End 12/2010
 
Description FEBS long-term fellowship (Anandapadamanaban)
Amount £29,010 (GBP)
Organisation Federation of European Biochemical Societies (FEBS) 
Sector Charity/Non Profit
Country European Union (EU)
Start 02/2017 
End 01/2018
 
Description Intellikine Inc
Amount £3,000 (GBP)
Organisation Intellikine, Inc. 
Sector Private
Country United States
Start 09/2008 
End 09/2009
 
Description LMB Cambridge Scholarship (Baretic) The Cambridge Trust
Amount £95,000 (GBP)
Organisation Medical Research Council (MRC) 
Department MRC Laboratory of Molecular Biology (LMB)
Sector Public
Country United Kingdom
Start 10/2010 
End 09/2013
 
Description LMB Cambridge Scholarship (Zhang) The Cambridge Trust
Amount £95,000 (GBP)
Organisation Medical Research Council (MRC) 
Department MRC Laboratory of Molecular Biology (LMB)
Sector Public
Country United Kingdom
Start 10/2009 
End 09/2012
 
Description MRC/AZ Cryo EM Structures and function studies of full length ATM & ATR: A molecular understanding of the regulation of DNA damage repair.
Amount £296,249 (GBP)
Funding ID BSF28 
Organisation AstraZeneca 
Sector Private
Country United Kingdom
Start 09/2017 
End 09/2020
 
Description MRC/AZ How do small-molecule activators of the PI3K superfamily work?
Amount £200,000 (GBP)
Funding ID BSF33 
Organisation AstraZeneca 
Sector Private
Country United Kingdom
Start 09/2018 
End 09/2020
 
Description Marie Curie Intra European Fellowship (Vadas)
Amount £124,296 (GBP)
Organisation Marie Sklodowska-Curie Actions 
Sector Academic/University
Country Global
Start 01/2012 
End 12/2012
 
Description Postdoc Fellowships for Research Abroad - Bioscience and Basic Biomedicine
Amount kr 3,440,571 (DKK)
Funding ID NNF18OC0030544 
Organisation Novo Nordisk Foundation 
Sector Charity/Non Profit
Country Denmark
Start 11/2018 
End 10/2022
 
Description Research Grant Intellikine
Amount $50,000 (USD)
Organisation Intellikine, Inc. 
Sector Private
Country United States
Start 12/2010 
End 09/2013
 
Description Research Grant Serono
Amount £281,240 (GBP)
Funding ID LSF 24463-blsf8555 
Organisation Merck 
Department Merck Serono
Sector Private
Country Germany
Start 11/2005 
End 10/2009
 
Description SNSF Fellowship (Vadas)
Amount £38,000 (GBP)
Organisation Swiss National Science Foundation 
Sector Public
Country Switzerland
Start 01/2011 
End 12/2011
 
Description SNSF Postdoctoral Fellowship (Vadas)
Amount £38,000 (GBP)
Organisation Swiss National Science Foundation 
Sector Public
Country Switzerland
Start 09/2009 
End 08/2010
 
Description St Catharines College JRF (Masson)
Amount £75,000 (GBP)
Organisation University of Cambridge 
Department St Catharine's College
Sector Academic/University
Country United Kingdom
Start 10/2015 
End 09/2018
 
Description Wellcome Trust Programme Grant (Assembly and disassembly ESCRTs on membranes)
Amount £561,453 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2008 
End 03/2013
 
Title PI3K crystallisation protocols 
Description We have developed detailed protocols for producing crystals of phosphoinositide 3-kinase gamma that can be used to determine the structures of complexes of the enzyme with small-molecule inhibitors. These include for expression in bacullovirus, purification, crystallisation, soaking crystals with compounds and crystal freezing protocols. We have communicated these protocols to various companies who have received materials from us. Without these protocols, it is difficult to produce high resolution structures. 
Type Of Material Technology assay or reagent 
Year Produced 2006 
Provided To Others? Yes  
Impact The protocols and the reagents that we have provided (viruses, purified protein and bacmids). Have facilitated many drug discovery programmes aimed at specific PI 3-kinase inhibitors. 
 
Title baculovirus expression system for IA PI3Ks 
Description We developed a baculovirus expression system that has enabled us to express lare quantities of class IA phhosphoinositide 3-kinases. The system has enabled us to express all of the mammalian PI 3-kinases in a manner that is amenable to determining high-resolution structures of the complexes of the enzyme with isotype-specific inhibitors. The system is particularly well suited to inhibitors of p110delta, p110alpha and p110beta. The details of this system have only ben disclosed at this time to our industrial collaborators at Merck-Serono. We expect to make full public disclosure by the end of 2008. 
Type Of Material Technology assay or reagent 
Year Produced 2008 
Provided To Others? Yes  
Impact This has allowed us to obtain the first structures of type IA PI 30kinases that are suitable for drug discovery programmes. 
 
Title co-expresson vectors for protein complexes 
Description We have developed a bacterial poly-cistronic expression system that we use to simultaneously express up to eight proteins. The system is used to express multi-subunit complexes with a variety of tags to facilitate X-ray crystallography, fluorescence spectroscopy and other biophysical techniques. The system involves a set of "shuttle" vectors as well as co-expression vectors. It has been been the basis for many of the structures that we have determined in the last five years, including the ESCRT complexes and catalytic/regulatory complexes for the PI 3-kinases. Work using these vectors has been published and the vectors have been provided to many researchers in the LMB and elsewhere. 
Type Of Material Technology assay or reagent 
Year Produced 2007 
Provided To Others? Yes  
Impact Without these vectors, many complexes would have been impossible to obtain. It is common that components of biological complexes cannot be obtained in isolation. These vectors have enabled us to go from no measurable expression for isolated components to massive yields of complexes suitable for X-ray crystallography. 
 
Description Bioinformatics of ESCRT complexes 
Organisation Miltenyi Biotec GmBH
Country Germany 
Sector Private 
PI Contribution We have determined the structure of ESCRT-containing complexes
Collaborator Contribution Kay Hoffman identified a novel ESCRT-I subunit
Impact One publication, pubmed ID 22405001
Start Year 2012
 
Description Biological inhibitors of cancers 
Organisation University of Leeds
Country United Kingdom 
Sector Academic/University 
PI Contribution We have assisted in determining structures of complexes of intracellular antibodies with the small G-protein Ras
Collaborator Contribution Prof. Rabbitts has explored the use of biological inhibitors such as intracellular antibodies for inhibiting cancers.
Impact Publication 17568777
 
Description Chemistry of PI3K inhibiors 
Organisation University of California, San Francisco
Country United States 
Sector Academic/University 
PI Contribution We determined the structures od a range of complexes of PI3Ks with isotype-selective inhibitors
Collaborator Contribution Colleages in Kevan Shokat's group have provided a wide range of compounds that are inhibitors of PI3Ks. These have been central to our ongoing structural studies of these enzymes.
Impact These collaborations have resulted in sev eral publications including: 16647110 18849971
 
Description Cryo-EM of ATM and ATR complexes 
Organisation Medical Research Council (MRC)
Country United Kingdom 
Sector Academic/University 
PI Contribution We are determining structures of ATM and ATR-containing complexes
Collaborator Contribution Sjors is providing guidance for analysing cryo-EM data
Impact No reportable outputs yet
Start Year 2015
 
Description Cryo-EM structures and functional studies of full length ATM & ATR: A molecular understanding of the regulation of DNA damage repair 
Organisation AstraZeneca
Department Research and Development AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We are determining structures of the PI3K-related protein kinases ATM and ATR.
Collaborator Contribution Through the AZ/LMB Blue Skies Innitiative, we have received funding for one postdoctoral researcher. AZ has also provided us large-scale cell cultures expressing the enzymes.
Impact There have been no reportable outcomes yet
Start Year 2015
 
Description Cryo-electron microscopy of a TOR complex 
Organisation Medical Research Council (MRC)
Country United Kingdom 
Sector Academic/University 
PI Contribution We have produced large quantities of pure complexes of the protein kinase TOR in a complex with LST8
Collaborator Contribution Dr. Vinothkumar has provided advice on cryo-EM, collected and analysed initial data sets and helped train members of my group
Impact One publication in press
Start Year 2015
 
Description Crystal structure of PI3K gamma 
Organisation Pfizer Ltd
Department Warner-Lambert
Country United States 
Sector Private 
PI Contribution We determined the first structure of a phosphoinositide 3-kinase.
Collaborator Contribution We collaborated in developing approches to designing PI3K inhibitors
Impact Protocols established in this collaboration resulted in a series of publications including pubmed ids: 10580505 11090628 11136978 16647110 18849971
 
Description Developing activators of lipid kinases 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We developed an assay to select for small molecules that would activate lipid kinases. We also will investigate the mechanism of activation using HDX-MS.
Collaborator Contribution AstraZeneca carried out a large-scale screen of a compound library using our assays. Our collaborators at UCL will be involved in using these inhibitors in various biological contexts.
Impact The collaboration has so far discovered a set of putative activators.
Start Year 2015
 
Description Developing activators of lipid kinases 
Organisation University College London
Department UCL Cancer Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution We developed an assay to select for small molecules that would activate lipid kinases. We also will investigate the mechanism of activation using HDX-MS.
Collaborator Contribution AstraZeneca carried out a large-scale screen of a compound library using our assays. Our collaborators at UCL will be involved in using these inhibitors in various biological contexts.
Impact The collaboration has so far discovered a set of putative activators.
Start Year 2015
 
Description Development of isotype-specific PI3K inhibitors 
Organisation University of California, San Francisco
Department Helen Diller Family Comprehensive Cancer Center
Country United States 
Sector Academic/University 
PI Contribution We have provided structures of complexes of PI3Ks with inhibitors
Collaborator Contribution Kevsn Shokat has provided isotype-spefic inhibitors
Impact Seven publications, PUBMED IDS: 16647110, 18849971, 20081827, 20154668, 20339072, 24876499, 26756197
 
Description Establishment of a PI3K structure-based drug develpment programme 
Organisation Vertex Pharmaceuticals
Country United States 
Sector Private 
PI Contribution We provided expression, purification and crystallisation protocols for PI3K gamma. We also provided plasmids and viruses for insect cell expression.
Impact Vertex established a structural component of their drug discovery programme for PI3K
 
Description Establishment of a structure-based PI3K drug discovery programme at Chiron 
Organisation Novartis
Department Chiron Corp
Country United States 
Sector Private 
PI Contribution We provided protocols for expression, purification and crystallisatiobn of PI3K gamma. We provided plasmids and viruses to facilitate expression.
Impact Chron established a PI3K structure programme for developing their inhibitors.
 
Description Establishment of a structure-based PI3K drug discovery programme at Sanofi-Aventis 
Organisation Sanofi
Department Aventis
Country France 
Sector Private 
PI Contribution We provided protocols for expression, purification and crystallisatiobn of PI3K gamma. We provided plasmids and viruses to facilitate expression.
Impact Sanofi-Aventis established a PI3K structure programme for developing their inhibitors.
 
Description HDX-MS studies of lipid transfer proteins 
Organisation Texas A&M University
Country United States 
Sector Academic/University 
PI Contribution We have analysed PI transfer proteins using HDX-MS to determine how they interact with lipids and membranes
Collaborator Contribution They have provided purified proteins
Impact No reportable output yet
Start Year 2015
 
Description HDX-MS to probe enzyme regulation and develop novel inhibitors of PI3Ks 
Organisation AstraZeneca
Department Research and Development AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We are using HDX-MS methods to develop inhibitors that prevent activation of PI3K complexes on membranes. In order to optimise this process, we are employing new MS fragmentation techniques.
Collaborator Contribution We meet regularly with our partners to discuss HDX-MS data acquisition and processing. We have participated in training postdocs in our partner organisation.
Impact There have not been any reportable outcomes yet
Start Year 2015
 
Description Inhibition of oncogenic PI3Ks 
Organisation Albert Einstein College of Medicine
Country United States 
Sector Academic/University 
PI Contribution We have determined the structures of class IA PI3Ks and used these structures to define mechanisms of oncogenesis by smatic mutations in these enzymes
Collaborator Contribution Prof. Backer has provided valuable expertise and reagents for expressing and characterising PI3Ks
Impact Publication 17626883
 
Description Mechanisms of PI3K-driven oncogenesis 
Organisation Albert Einstein College of Medicine
Country United States 
Sector Academic/University 
PI Contribution We have analysed regulation of PI3K complexes using HDX-MS
Collaborator Contribution Jonathan has examind the impact of input-ablated mutants on oncogenesis in vitro
Impact Four publications, PUBMED IDs, 17626883, 23211529, 23328076, 24190998
Start Year 2011
 
Description Modelling flexibility in PI3Ks 
Organisation Intellikine, Inc.
Country United States 
Sector Private 
PI Contribution We determined structures of complexes of isotype-specific inhibitors of PI3K delta with specific inhbitors developed by Intellikine. We also developed computational tools to model the flexibility of PI3Ks.
Collaborator Contribution We have had ongoing discussions related to designing new PI3K inhibitors
Impact We have published our structures and conclusions in Nature Chemical Biology. This is in press and due to be published in February, 2010.
 
Description Nanobodies for structural studies 
Organisation Flemish Institute for Biotechnology
Country Belgium 
Sector Academic/University 
PI Contribution We have provided cross-linked PI3K-containing complexes for immunization
Collaborator Contribution Jan Steyaert has provided nanobodies that bind to Vps34-containing complexes and complexes of a class IA PI3K
Impact One publication, 26450213
Start Year 2012
 
Description PI3K delta activity in pathogenesis of APDS 
Organisation University of Cambridge
Department Department of Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution We have characterised the activity and membrane interqactions of mutants of p110 delta associated with familial immune disorders
Collaborator Contribution Alison has characterised the cells of APDS patients
Impact One publication, pubmed id 24136356
Start Year 2012
 
Description Phosphoinositide recognition moduels 
Organisation University of Illinois
Country United States 
Sector Academic/University 
PI Contribution We have determined the structures of phosphoinositide-recognition modules such as PX domains
Collaborator Contribution Prof. Cho has characterised the physics of interactions of recognition modules for lipid membranes. This has helped us understand our structures of these modules.
Impact Publications: 17038310 17581820 1764168
 
Description Protein trafficking to lysosomes via ESCRT complexes 
Organisation Cornell University
Country United States 
Sector Academic/University 
PI Contribution We have determined three-dimensional structures of a range of endosomal complexes involved in sorting membrane proteins into multivesicular bodies for degradation in lsosomes
Collaborator Contribution We have published a series of papers with our structures and cellular sorting results from Scot Emr's groupDr. Urbe has been helped us decide important research directions that understanding lysosomal trafficking
Impact Our structures have been the basis for several publications 16615893 17215868 17450176
 
Description Protein trafficking to lysosomes via ESCRT complexes 
Organisation University of Liverpool
Country United Kingdom 
Sector Academic/University 
PI Contribution We have determined three-dimensional structures of a range of endosomal complexes involved in sorting membrane proteins into multivesicular bodies for degradation in lsosomes
Collaborator Contribution We have published a series of papers with our structures and cellular sorting results from Scot Emr's groupDr. Urbe has been helped us decide important research directions that understanding lysosomal trafficking
Impact Our structures have been the basis for several publications 16615893 17215868 17450176
 
Description Quantitative analysis of the effects of membrane curvatute on activity of Vps34 complexes 
Organisation University of Geneva
Department Department of Biochemistry
Country Switzerland 
Sector Academic/University 
PI Contribution We have provided purified Vps34 complexes and assayed them as a function of vesicle size and lipid composition
Collaborator Contribution They have drawn lipid tubules to generate high-curvature structures on which to assay the Vps34 complexes
Impact No reportable output yet
Start Year 2015
 
Description Regulation of autophagy induction by the VPS34 complexes: structural and functional studies 
Organisation Babraham Institute
Department Signalling
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution We have determined structures, in vitro activities, dynamics and membrane binding for several Vps34-containing complexes
Collaborator Contribution Nicholas has assayed autophagy in mammalian cells
Impact One publication, PUBMED ID 26450213
Start Year 2012
 
Description Regulation of autophagy induction by the Vps34 complex: structural and functional studies 
Organisation Babraham Institute
Department Signalling
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution We are collaborating on the structures of Vps34-containing complexes
Collaborator Contribution Our work is primarily structural. The structures we are determining are being used as a framework for designing functional studies of these complexes by researchers in the Babraham Institute.
Impact Several structures have been determined and publictions are in progress
Start Year 2012
 
Description Regulation of class IB PI3Ks 
Organisation Eberhard Karls University of Tubingen
Country Germany 
Sector Academic/University 
PI Contribution We have carried HDX-MS studies of class IB PI3Ks and determined their activities in vitro
Collaborator Contribution They have provided lipidated Gbeta/gamma subunits expressed in SF9 cells
Impact Three publications, PUBMED IDS: 3211529, 24190998 and 26173259
Start Year 2011
 
Description Role of ESCRTs in archaea 
Organisation University of Oxford
Department Medical Sciences Division
Country United Kingdom 
Sector Academic/University 
PI Contribution We have determined the structures of complexes of ESCRT-replated proteins from archaeal cells
Collaborator Contribution Prof. Bell has been essential in determining the roles of ESCRT-related proteins in archaeal cells
Impact Publication 19008417
 
Description Single-molecule imaging of PI3K and PTEN activity on membranes 
Organisation University of Colorado Boulder
Department Department of Chemistry and Biochemistry
Country United States 
Sector Academic/University 
PI Contribution We have provided large quantities of purified PI3K and PTEN
Collaborator Contribution Joseph has characterised the activity, diffusion and lipid interactions using single-molecule spectrometry
Impact Two publications
Start Year 2012
 
Description Structural and functional studies of PI3Ks 
Organisation Babraham Institute
Department Signalling
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution We have determined structures of PI3K-containing complexes and analysed their dynamics by HDX-MS. We have also characterised in vitro activitiies of these complexes
Collaborator Contribution They have assayed PI3K complexes in cells and mice
Impact Six publications, PUBMED IDS: 11090628 11136978 11684018 15581496 21362552 24136356
 
Description Structural and functional studies of PI3Ks 
Organisation University of Basel
Country Switzerland 
Sector Academic/University 
PI Contribution We have carried out HDX-MS studies of p110gamma/p84 complexes
Collaborator Contribution Matthis Wymann has determine the activity of PI3K gamma complexes in cells
Impact publication 23824069
 
Description Structural studies of golgi-associated proteins 
Organisation Medical Research Council (MRC)
Country United Kingdom 
Sector Academic/University 
PI Contribution We have helped researchers in Sean Munro's lab to determine structures of golgi-associated proteins.
Collaborator Contribution They have helped us with assays of intracellular protein sorting.
Impact One publication, PUBMED ID 14580338, Another in review.
 
Description Structure and function of MITD1 
Organisation Guy's Hospital
Country United Kingdom 
Sector Hospitals 
PI Contribution We have determined structures of MITD1 and associated proteins
Collaborator Contribution Monica has determined the role of MITD1 in cells
Impact Two publications, PUBMED ID 23045692 and 22405001
Start Year 2012
 
Description Structure and function of phosphoinositide kinases 
Organisation University of Cambridge
Department Department of Pharmacology
Country United Kingdom 
Sector Academic/University 
PI Contribution We have analysed the dynamics and membrane binding of phosphtidylinositol kinases, using HDX-MS
Collaborator Contribution They have provided the ptoein complexes for analysis
Impact Two publications: 15350214, 25495341
 
Description Structure-based drug discovery for PI3K delta inhibitors 
Organisation Merck
Department Merck Serono
Country Germany 
Sector Private 
PI Contribution We established protocols for expressing and purifying large quantities of phosphoinositide 3-kinase alpha and delta for structure-based drug discovery programmes. We crystallised and determned structures of a wide ranger of complexes of these enzymes with inhibitors designed by Serono and others. Using these structures, we established mechanisms that underpin one class of isotype-specific inhibitor.
Collaborator Contribution We have extensive, ongoing discussions regarding important directions in designing PI3K inhbitors
Impact We published our conclusions regarding isotype-specific inhibitors of PI3K delta in Nature Chemical Biology. This is currently in press and tentatively scheduled for publication in February, 2010.
 
Description Structures of helical assemblies by electron microscopy 
Organisation University of Virginia (UVa)
Department Health Sciences Centre
Country United States 
Sector Academic/University 
PI Contribution We deterines the structure of ESCRT-III filaments by electron microscopy with the help of Prof. Egelman
Collaborator Contribution Prof. Egelman provided expertise necessary to determine the structures of helical assemblies of ESCRT complexes
Impact Publication 18786397
 
Description Structures of spindle-pole body complexes 
Organisation Medical Research Council (MRC)
Department MRC Laboratory of Molecular Biology (LMB)
Country United Kingdom 
Sector Public 
PI Contribution We have helped determine protein structures of complexes involved in spindle-pole bodies
Collaborator Contribution They have provided crystals of purified protein complexes
Impact We have published the structure and the invivo assembly of components of the spindle-pole body pubmed id: 16785321
 
Description Studies of phosphoinositide signalling using single molecule fluorescence methods on supported bilayers 
Organisation University of Colorado Boulder
Department Department of Chemistry and Biochemistry
Country United States 
Sector Academic/University 
PI Contribution We have provided samples of various purified enzymes for studies of activity and difussion on membranes
Collaborator Contribution Falke and his clleagues have used our material to characterise the molecular mechanisms whereby the activity of the enzymes is regulate on membranes
Impact Two publications: PMID: 27933776 PMID: 27119641
Start Year 2012
 
Description The molecular basis of PI3Kgamma in autoimmune lymphoproliferative syndrome 
Organisation National Institute of Allergy and Infectious Diseases (NIAID)
Country United States 
Sector Public 
PI Contribution We have assayed the PI3K activity of p110gamma mutants associated with familial autoimmune disease
Collaborator Contribution Our partners have assayed the inluence of the p110gamma mutations in human cells
Impact No reportable output yet
Start Year 2015
 
Description The molecular basis of SHORT syndrome 
Organisation University of Cambridge
Department Institute of Metabolic Science (IMS)
Country United Kingdom 
Sector Academic/University 
PI Contribution We have provided p110alpha complexes and trained a Ph.D. student in work with mutants of the enzyme
Collaborator Contribution The Ph.D. student has cloned SHORT syndrome mutants, expressed them and assayed their activity
Impact No reprtable output yet
Start Year 2014
 
Description The role of PTEN in Wnt signalling 
Organisation Mount Sinai Hospital (Canada)
Department Samuel Lunenfeld Research Institute
Country Canada 
Sector Hospitals 
PI Contribution We assayed in vitro activities of PTEN mutants
Collaborator Contribution They characterised the role of PTEN in cilia formation
Impact One publication, 26399523
Start Year 2014
 
Description The structure of PI3K alpha 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We expressed, purified and crystallised the core of the p110 alpha/p85 complex. We determined the structure of the adaptor-binding domain of p110 alpha in a complex with the iSH2 domain from p85 regulatory subunit.
Collaborator Contribution Over the period of the collaboration, we have discussed directions important for development of PI3K inhibitors
Impact This resulted in the publication f the structure and our conclusions in the journal Science. Pubmed id 17626883
 
Description structure and function of p110 beta/ p85 beta 
Organisation Babraham Institute
Country United Kingdom 
Sector Private 
PI Contribution We have determined structures of the complexes of p110 beta with regulatory subunits
Collaborator Contribution They have been involved in various aspects of our reserch programme, including mass spectrometry for assaying PI3Ks and mouse studies of structure guided mutations
Impact publication 21362552
Start Year 2010
 
Description Big Biology Day 2108. Synthetic biology: Create a New Protein! 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Yohei Ohashi was a member of a team that visited Hills Road 6th Form to interest students in science. His focus was "Synthetic biology: Create a New Protein!"
Year(s) Of Engagement Activity 2018
 
Description Biochemical Society Big Biology Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Two members of the group participated in a "Medicine Makers" event developed by the Biochemical Society and British Pharmacological Society. This was held in Cambridge at the at "Big Biology Day".
Year(s) Of Engagement Activity 2016
 
Description Cambridge Science Festival 2018. Worms are cleverer than you think! 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Yohei Ohashi helped explain how C. elegans is used as a model research organism
Year(s) Of Engagement Activity 2018
 
Description FindAPhD Fair - Cambridge 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact Ksenia Rostislavleva, a Ph.D. student in the group, helped assemble a set of posters and a stand to advise undergratuates about careeres in research at the Laboratory of Molecular Biology
Year(s) Of Engagement Activity 2015
 
Description Home in the Service of Science 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact "Home in the Service of Science" is an art installation in the LMB. Olga Perisic in the group participated in a public event to open the installation for the general public.
Year(s) Of Engagement Activity 2016
 
Description Long Road 6th form 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact About 10 sixth form students toured the lab, and we answered questions about research and medical careers
Year(s) Of Engagement Activity 2014
 
Description Ridgefield Primary School Science Day 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Presentation for school children by Glenn Masson
Year(s) Of Engagement Activity 2012
 
Description Royal Society annual exhibition 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Olga Perisic spent a day as a volunteer talking to visitors at a stand at the Royal Society annual exhibition called "Virus wars - antibodies strike back" which celebrated our immune system, research and 100 years of the MRC , Saturday 6th July 2013.
Year(s) Of Engagement Activity 2013
 
Description Science Festival, Cambridge 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Glenn Masson helped man am LMB booth at the Science Festival
Year(s) Of Engagement Activity 2015
 
Description Structural biology as a tool for finding new pharmaceutical agents 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact The group of sixth form students who visited the institute learned about the mission of the LMB and some basic principles of strutural biology as applied to drug discovery. In addition, they took part in a discussion with several scientists on various aspects of a career as a scintist and practical information for applying to University,

After the talk in 2013, the sixth form requested that we repeat this is 2014.
Year(s) Of Engagement Activity 2013,2014
 
Description long road 6th form 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach Local
Primary Audience Schools
Results and Impact We hosted the visit of 10 top 6th form students to talk about careers in molecular biology with an emphasis on molecular medicine.

The head of studies at the sixth form reported that students found the visit helpful and that they were interested in establishing a work experience program.
Year(s) Of Engagement Activity 2013