Upload
kendall-yelverton
View
227
Download
0
Tags:
Embed Size (px)
Citation preview
BBSRC David Phillips Research Fellowship an interdisciplinary perspective
Dr. Ed Tate, Department of Chemistry, ICL
• Context:• Current research• Career
• Fellowship Applications:• Personal perspective • Hints and Tips
• Research fellowship:• Advantages, challenges
Faculty of Medicine Fellowships DayJuly 2008
Research interests
1. Development & application of new technologies in chemical biology New chemical approaches for site-specific protein labelling Chemical tools for exploring post-translational modification of proteins Reactive probes for in vitro & in vivo enzyme activity profiling & imaging
2. Design and synthesis of small molecule inhibitors of: Protein-protein interactions: site-specific disruption of multi-protein complexes
• Martin Buck (CBC), Xiaodong Zhang (CBC), Tony Holder (NIMR) Protein-ligand interactions: transferase and protease inhibitors
• Neil Fairweather (CMMI), Miguel Seabra (NHLI), Debbie Smith (York)
Dr. Ed Tate, Department of ChemistryBBSRC David Phillips Research Fellow (since 2006)
[email protected]; http://www3.imperial.ac.uk/people/e.tate/research
Chemical proteomics Applied organic chemistry of living systems Targets complex post-translational
phenomena: PTMs, enzyme activity in living systems, protein-protein interactions…
Proteomic complement of chemical genetics Protein labelling, imaging, biomarker
analysis…
What is chemical proteomics?
DNA RNA Protein
Transcription Translation
Post-translational modification
>300 types: Phosphorylation, glycosylation, lipidation, acetylation, proteolysis…
Origin
Post-translational Modifications
Phosphorylation
Signalling
Glycosylation
Immune system
Cell adhesion
Prenylation
Trafficking
Acylation
Membrane association
Acetylation
Transcriptional regulation
Proteolysis
Apoptotic cascade
Ubiquitination
Protein degradation
Site Dynamics
Function
Chemical Proteomics
Sulfenic acids
Redox response
Site-specific labelling of proteins
Metabolic tagging
Target proteins tagged at site of PTM
Protein bearing site-specific chemical feature
(active site, PTM)
Tagged PTM applied to
cells/animal
Proteins labelled site-specifically
- + ++
Drug mode-of-action studies
ID proteins and site of PTM
Live- and fixed-cell imaging
Bioorthogonal ligation chemistry
Chemical or enzymatic tagging
A platform for Chemical Myristomics
(‘Azido-myristic’ acid)
N3
HN
O
N3
HN
O
N3
HN
O
N3
S
O
CoA
N3
OH
O
N3
OH
O
N3
HN
O
N
NN
HN
O
Secondary Labels
Bioorthogonal ligation
Secondary Labels
MudPIT analysis
2D-DIGEOn-bead purification
Activity-based probesSurface layer formation in C. difficile
C. difficile Spore-forming anaerobe Most lethal hospital superbug Resistant to most antibiotics Lack of genetic tools
Excretes a crystalline S-layer Post-translational cleavage Unknown cysteine protease
O
HOOC
O
HN LysTyrGlu PEG3 Biotin
WarheadSpecificity
element
Linker
LabelFeed to C. difficile Affinity purify, ID
Chemical Technologies New chemical labelling
technologies New chemistries Extend to other PTMs ABPs for new enzyme activities
Discovery of new PTMs Pull out after metabolic labelling
Nucleotide tagging Protein-DNA interactions Transcriptional activation
Future perspectives
Biomedical Applications
+ Chemical genetics Drug screening Biomarker analysis
+ Synthetic biology Introducing new chemistries
+ Live cell & in vivo imaging Trafficking due to PTM Cell-surface display
+ Systems biology Dissect pathways Probes for protein-protein &
protein-membrane interactions
Career OverviewPhD and Postdoctoral Work
BSc in Chemistry (Durham)
1993-
1996
PhD in Organic Chemistry(Cambridge)
1996-
1999
1851 Research Fellowship(Ecole Polytechnique)
2000-
2001
Howard Trust Research Fellowship (Institut Pasteur)
2002-
2003
BBSRC-funded PDRA (ICL)
2004-
2006
New C-glycosidation reactionsTotal synthesis of natural products
Radical cyclisation chemistryTotal synthesis of natural products
Role of DNA helix stability & upstream sequences in transcriptional regulation
Protein/peptide synthesis and engineeringLibrary generation and screening techniques
Application StageSituation as of August 2005
Prior Applications: WT fellowship application in 2005 rejected – insufficient
biochemical track record Never been involved in writing a standard grant application!
Personal Situation: Current PDRA contract due to expire 1 month after expected
fellowship decision, no follow-on funding… Baby due 22nd November!
Research Track Record: Good chemical research track record No biochem research track record at time of application
Application StagePlan of Action
Applied for multiple fellowships EPSRC Advanced Research Fellowship (ARF) BBSRC David Phillips Fellowship Royal Society URF Wellcome Trust MRC
Novel proposal: chemical proteomics Topical and interdisciplinary Very limited literature precedent Minimal preliminary data, and no prior track record!
Plan B: other grant proposals Co-I on MRC Discipline-Hopping grant (related subject) ‘Researcher Co-I’ on BBSRC responsive mode grant (different subject)
Application StageMaking Multiple Applications
Address Remit Direct your proposal towards the funder’s remit
BBSRC: basic biology, complete independence (PI), 3-year postdoc MRC: medical emphasis, strong training/career development component Wellcome: technician for 5 years, previously had to have HoD as PI EPSRC: steer clear of biological outputs (stick to technology only) Royal Society: two FRS’s strongly supporting application, no support grant.
Remit is strictly based on the research outputs (‘deliverables’) of your
project (not the methodology and techniques applied)
Be flexible, rewrite application to take account of funding available, in
particular in justifying your costs
Advantages Spreads the risk of hitting a referee with strong negative bias Not always clear in advance of submission which funder is most
appropriate
Application StageWriting Applications
Writing the proposal Steep learning curve, very time-consuming (2-3 months) Sole PI, so responsible for all aspects of the proposal: financial,
planning, scientific Very good practice for later PI grant applications!
Costing the proposal Costing may feel rather abstract on the application, but you will
appreciate careful costing if you get the fellowship. Acquire a working knowledge of Full Economic Costing (fEC) The headline cost can be big: >£1 million for BBSRC DP Fellowship –
but 30%+ goes direct to Faculty under fEC… Request costs at the upper limit allowed if you can justify them for the
proposed work. You can usually move cash around later (within limits!)
Application StageGeneral Tips
Create a career narrative Justify your career choices (in hindsight) Highlight why you are ready to go independent
Fellowship proposals vs. grant proposals An original, cutting-edge idea can carry more weight than a strong
track record (esp. when compared to standard grants) You can get away with having (much) less preliminary data Interdisciplinary research may fare better in fellowship
applications than in standard grant applications.
Salary level Most funders don’t fix the level arbitrarily You don’t usually need to justify your own salary level
Some more things to think about… Scientific merit – external peer review (it is a grant proposal!) Address important questions – originality, fresh thinking Has to be feasible – you have to convince the referees that you
are capable of doing the work proposed, in the time available Show them that you have considered the riskier elements, and
propose contingency plans; don’t have the whole project dependent on a risky proposition
Mention preliminary data if you have it – I would say this is not absolutely essential, but you do need to be able to convince the referees that the riskier aspects could work
Know your competition in the scientific field – cite the key work of others in your background section
Cheaper proposals do not mean more chance of success; but high-cost proposals where costs are not justified will also not impress.
Application StageWriting Applications
Application StageStay or Go?
• Stay… Access to existing equipment and collaborations Minimal start-up time Can you achieve independence?
• In either case…• Ensure your expectations (support, space, teaching etc.) are understood
by host institute• Be prepared to justify choice of institute in the application and at interview• Be prepared for questions regarding future independence if you decide to
stay
• Or go? Start afresh on your own terms Will take time to get started• May help to get out of the ‘golden triangle’…
Interview StageSome suggestions
Presentation Practise in front of a diverse audience Keep it simple!
Interview BBSRC interview is very brief (20min), and (relatively) friendly. Interviews for other funders can be more intense… Be prepared for questions on: Past career choices How will a fellowship benefit you and your research? Minimal teaching, independence, opportunity to apply for
further grants during fellowship. How will you achieve independence? Where do you see yourself in 5 years, 10 years… Permanent position in academia, leader of a vibrant research
group
Award StageChallenges and Benefits
Benefits Minimal teaching load (vs. lectureship) = Time to set up collaborations, supervise research, write new grants… Support from BBSRC Once in post, be pro-active in applying for further funding:
From October: 14 (3 PDRAs, 9 PhD + 2 UG research students) Over £1 million in PI research income (+ £1 million fellowship)
Challenges Making the transition from PDRA to PI
Managing & applying for grants, recruitment Supervising a group on your own Dealing with internal departmental politics
You only have a few years to find a permanent position! Start looking after 1-2 years May entail a move to another institute Consider whether fellows have routinely been taken on permanently at the
host department/institute
Acknowledgements
JemimaGillianTamTook
PinarAlexLucy
Postgraduate Students (PhD and MRes)
Will Sasala
Postdoctoral Researchers
James Camille
Undergraduate Students