Contact: Astbury 10.116, +44(0) 113 34 37821,
You can read more about Dr Brockwell's interests here:
The effects of force on proteins and their complexes; extremophilic proteins; membrane protein folding and folding factors.
Currently research in the group is following three themes which are described below. If you are interested in joining the lab to study for a PhD find studentship and eligibility details here.
Our Asylum Research AFM instruments are housed in the School of Physics and Astronomy at Leeds. This allows regular contact with colleagues Simon Connell, Lorna Dougan and Neil Thomson. Areas from which our research benefits from the input of colleagues in Leeds include the Structural Molecular Biology and Integrative Membrane Biology groups.
The effects of force on proteins and their complexes
Mechanical unfolding: in vitro many proteins are required to resist or respond to mechanical stimuli. Over the last decade the development of atomic force microscope (AFM) instruments with high force sensitivity and sub-nanometre distance resolution has allowed the mechanical properties of single protein molecules to be measured. This work has revealed that proteins with similar stability to chemical denaturants can behave very differently when unfolded by the AFM. We are interested in finding out why some proteins are able to resist greater unfolding forces than others to understand how Nature uses mechanical denaturation to accelerate protein unfolding. In addition, we also use this information to rationally design proteins with novel mechanical properties.
Publication: Identification of a mechanical rheostat in the hydrophobic core of protein L. Sadler et al. (2009) J Mol Biol 393:237-248.
Exploring protein-ligand interactions: in addition to characterising the behaviour of proteins under mechanical extension we have also started to measure the mechanical strength of the non-covalent interactions between proteins and their ligands. In particular we are interested in finding out how very strong interactions are broken apart on timescales fast enough to be biochemically useful. To do this we specifically immobilise one protein partner onto a surface and the other onto the tip of the AFM cantilever. This work has revealed that highly avid complexes can be rapidly broken apart by the application of small forces that are accessible to Nature.
Publication: A force-activated trip switch triggers rapid dissociation of a colicin from its immunity protein. Farrance et al. (2013) PLoS Biol 11:e1001489.
Properties of extremophilic proteins
Life is found in the harshest of environments: low and high temperatures (< 0 and > 100 °C, respectively), low pH (< pH 2) and high salt concentrations (> 2M). For many uni-cellular organisms, adaption to survival in these environments often occurs by the evolution of extremophilic proteins e.g. those that remain functional at low or high temperatures (psychrophilic and hyperthermophilic proteins). In a collaboration with Lorna Dougan (Physics, Leeds), we have recently started to examine the biophysical and mechanical properties of a range of extremophilic proteins.
Publication: Single-molecule force spectroscopy identifies a small cold shock protein as being mechanically robust. Hoffmann et al. (2013) J Phys Chem B 117:1819−1826.
Membrane protein folding and folding factors
Despite their ubiquity and importance as cellular gatekeepers, progress in understanding how membrane proteins fold into the narrow ensemble of structures required for their function is slow. In a collaboration with Professors Steve Baldwin and Sheena Radford we are examining the folding and insertion of the bacterial outer membrane protein PagP into liposomes and how periplasmic chaperones facilitate this process.
Publication: Malleability of the folding mechanism of the outer membrane protein PagP: parallel pathways and the effect of membrane elasticity. Huysmans et al. J Mol Biol 416:453-463.
BIOC1303 - Introductory Biochemistry: Problem Solving and Data Handling
BIOC1301 - Introductory Integrated Biochemistry: the Molecules and Processes of Life
BIOC1302 - Introductory Biochemistry: Practical Skills
BIOC1303 - Introductory Biochemistry: Problem Solving and Data Handling
BIOC2301 - Intermediate Integrated Biochemistry
BIOC2302 - Intermediate Biochemistry: Practicals
BIOC2303 - Intermediate Biochemistry: Skills
BIOC3160 - Laboratory/Literature/Computing Research Project
BIOC3221/22 b - ATU - Protein Dynamics
BIOC3221/22/BIOL3210 b - ATU - Folding & Diseases
BIOL1112 - The Molecules of Life
BIOL3399 - Extended Research Project Preparation
BIOL5380M - Extended Research Project
BMSC2120 - Scientific Skills
Member of Masters Taught Student Education Committee (Exams Officer: Bioscience)Member of Undergraduate School Taught Student Education Committee (Exams Officer: Biochemistry, Microbiology & Biological Sciences)
Centre membership: The Astbury Centre for Structural Molecular Biology
Ciaran Doherty (Primary supervisor) 65% FTE
Samuel Hickman (Primary supervisor) 34% FTE
Rhys Thomas (Primary supervisor) 50% FTE
Leon Willis (Primary supervisor) 34% FTE
John Dobson (Co-supervisor) 50% FTE
Anna Higgins (Co-supervisor) 50% FTE
James Horne (Co-supervisor) 50% FTE
Julia Humes (Co-supervisor) 50% FTE
Katherine Kendrick (Co-supervisor) 20% FTE
Robert Schiffrin (Co-supervisor) 50% FTE
Michael Wilson (Co-supervisor) 50% FTE
Schiffrin B, Calabrese AN, Devine PWA, Harris SA, Ashcroft AE, Brockwell DJ, Radford SE Skp is a multivalent chaperone of outer-membrane proteins Nature Structural and Molecular Biology 23 786-793, 2016
Tych KM, Batchelor M, Hoffmann T, Wilson MC, Hughes ML, Paci E, Brockwell DJ, Dougan L Differential Effects of Hydrophobic Core Packing Residues for Thermodynamic and Mechanical Stability of a Hyperthermophilic Protein Langmuir 32 7392-7402, 2016
Tych KM, Batchelor M, Hoffmann T, Wilson MC, Paci E, Brockwell DJ, Dougan L Tuning protein mechanics through an ionic cluster graft from an extremophilic protein Soft Matter 12 2688-2699, 2016
Saunders JC, Young LM, Mahood RA, Jackson MP, Revill CH, Foster RJ, Smith DA, Ashcroft AE, Brockwell DJ, Radford SE An in vivo platform for identifying inhibitors of protein aggregation Nature Chemical Biology 12 94-101, 2016
Hoffmann T, Tych KM, Crosskey T, Schiffrin B, Brockwell DJ, Dougan L Rapid and Robust Polyprotein Production Facilitates Single-Molecule Mechanical Characterization ofβ-Barrel Assembly Machinery Polypeptide Transport Associated Domains ACS Nano 9 8811-8821, 2015
Tych KM, Hoffmann T, Batchelor M, Hughes ML, Kendrick KE, Walsh DL, Wilson M, Brockwell DJ, Dougan L Life in extreme environments: single molecule force spectroscopy as a tool to explore proteins from extremophilic organisms. Biochem Soc Trans 43 179-185, 2015
Tych KM, Hughes ML, Bourke J, Taniguchi Y, Kawakami M, Brockwell DJ, Dougan L Optimizing the calculation of energy landscape parameters from single-molecule protein unfolding experiments Physical Review E - Statistical, Nonlinear, and Soft Matter Physics 91 -, 2015
Farrance OE, Paci E, Radford SE, Brockwell DJ Extraction of accurate biomolecular parameters from single-molecule force spectroscopy experiments ACS Nano 9 1315-1324, 2015
Gruszka DT, Whelan F, Farrance OE, Fung HK, Paci E, Jeffries CM, Svergun DI, Baldock C, Baumann CG, Brockwell DJ, Potts JR, Clarke J Cooperative folding of intrinsically disordered domains drives assembly of a strong elongated protein. Nat Commun 6 7271-, 2015
McMorran LM, Brockwell DJ, Radford SE Mechanistic studies of the biogenesis and folding of outer membrane proteins in vitro and in vivo: What have we learned to date? Archives of Biochemistry and Biophysics -, 2014
McMorran LM, Bartlett AL, Huysmans GHM, Radford SE, Brockwell DJ Dissecting the effects of periplasmic chaperones on the in vitro folding of the outer membrane protein PagP Journal of Molecular Biology 425 3178-3191, 2013
Hoffmann T, Tych KM, Brockwell DJ, Dougan L Single-molecule force spectroscopy identifies a small cold shock protein as being mechanically robust. J Phys Chem B 117 1819-1826, 2013
Farrance OE, Hann E, Kaminska R, Housden NG, Derrington SR, Kleanthous C, Radford SE, Brockwell DJ A force-activated trip switch triggers rapid dissociation of a colicin from its immunity protein. PLoS Biol 11 e1001489-, 2013
Hoffmann T, Tych KM, Hughes ML, Brockwell DJ, Dougan L Towards design principles for determining the mechanical stability of proteins Physical Chemistry Chemical Physics 15 15767-15780, 2013
Tych KM, Hoffmann T, Brockwell DJ, Dougan L Single molecule force spectroscopy reveals the temperature-dependent robustness and malleability of a hyperthermophilic protein Soft Matter 9 9016-9025, 2013
Ikeda-Kobayashi A, Taniguchi Y, Brockwell DJ, Paci E, Kawakami M Prying open single GroES Ring complexes by force reveals cooperativity across domains Biophysical Journal 102 1961-1968, 2012
Huysmans GH, Radford SE, Baldwin SA, Brockwell DJ Malleability of the folding mechanism of the outer membrane protein PagP: parallel pathways and the effect of membrane elasticity. J Mol Biol 416 453-464, 2012
Crampton N, Alzahrani K, Beddard GS, Connell SD, Brockwell DJ Mechanically Unfolding Protein L Using a Laser-Feedback-Controlled Cantilever BIOPHYS J 100 1800-1809, 2011
Crampton N, Brockwell DJ Unravelling the design principles for single protein mechanical strength CURR OPIN STRUC BIOL 20 508-517, 2010
Taniguchi Y, Khatri BS, Brockwell DJ, Paci E, Kawakami M Dynamics of the Coiled-Coil Unfolding Transition of Myosin Rod Probed by Dissipation Force Spectrum BIOPHYS J 99 257-262, 2010
Pugh SD, Gell C, Smith DA, Radford SE, Brockwell DJ Single-Molecule Studies of the Im7 Folding Landscape J MOL BIOL 398 132-145, 2010
Huysmans GHM, Baldwin SA, Brockwell DJ, Radford SE The transition state for folding of an outer membrane protein P NATL ACAD SCI USA 107 4099-4104, 2010
Beddard GS, Brockwell DJ A statistical approach to the estimation of mechanical unfolding parameters from the unfolding patterns of protein heteropolymers PHYS BIOL 7 -, 2010
Grant CA, Brockwell DJ, Radford SE, Thomson NH Tuning the Elastic Modulus of Hydrated Collagen Fibrils BIOPHYS J 97 2985-2992, 2009
Sadler DP, Petrik E, Taniguchi Y, Pullen JR, Kawakami M, Radford SE, Brockwell DJ Identification of a Mechanical Rheostat in the Hydrophobic Core of Protein L J MOL BIOL 393 237-248, 2009
Taniguchi Y, Brockwell DJ, Kawakami M The Effect of Temperature on Mechanical Resistance of the Native and Intermediate States of I27 BIOPHYS J 95 5296-5305, 2008
Grant CA, Brockwell DJ, Radford SE, Thomson NH Effects of hydration on the mechanical response of individual collagen fibrils APPL PHYS LETT 92 -, 2008
Anderson KL, Radford SE, Smith DA, Brockwell DJ The dynamical response of proteins under force In Handbook of molecular force spectroscopy , 2008
Khatri BS, Byrne K, Kawakami M, Brockwell DJ, Smith DA, Radford SE, McLeish TCB Internal friction of single polypeptide chains at high stretch FARADAY DISCUSS 139 35-51, 2008
Brockwell DJ Probing the mechanical stability of proteins using the atomic force microscope BIOCHEM SOC T 35 1564-1568, 2007
Huysmans GH, Radford SE, Brockwell DJ, Baldwin SA The N-terminal helix is a post-assembly clamp in the bacterial outer membrane protein PagP. J Mol Biol 373 529-540, 2007
Hann E, Kirkpatrick N, Kleanthous C, Smith DA, Radford SE, Brockwell DJ The effect of protein complexation on the mechanical stability of Im9 BIOPHYS J 92 L79-L81, 2007
Brockwell DJ Force denaturation of proteins - an unfolding story Current Nanoscience 3 3-15, 2007
Brockwell DJ, Radford SE Intermediates: ubiquitous species on folding energy landscapes? CURR OPIN STRUC BIOL 17 30-37, 2007
Tezuka-Kawakami T, Gell C, Brockwell DJ, Radford SE, Smith DAM Urea-Induced Unfolding of the Immunity Protein Im9 Monitored by spFRET Biophysical Journal 91 L42-L44, 2006
West DK, Brockwell DJ, Paci E Prediction of the translocation kinetics of a protein from its mechanical properties BIOPHYS J 91 L51-L53, 2006
Kawakami M, Byrne K, Brockwell DJ, Radford SE, Smith DA Viscoelastic study of the mechanical unfolding of a protein by AFM Biophysical Journal 91 L16-L18, 2006
West DK, Brockwell DJ, Olmsted PD, Radford SE, Paci E Mechanical resistance of proteins explained using simple molecular models BIOPHYS J 90 287-297, 2006
Brockwell DJ, Beddard GS, Paci E, West DK, Olmsted PD, Smith DA, Radford SE Mechanically unfolding the small, topologically simple protein L BIOPHYS J 89 506-519, 2005
Brockwell DJ, Paci E, Zinober RC, Beddard GS, Olmsted PD, Smith DA, Perham RN, Radford SE Pulling geometry defines the mechanical resistance of a beta-sheet protein. Nat Struct Biol 10 731-737, 2003
Best RB, Brockwell DJ, Toca-Herrera JL, Blake AW, Smith DA, Radford SE, Clarke J Force mode atomic force microscopy as a tool for protein folding studies ANAL CHIM ACTA 479 87-105, 2003
Smith DAM, Brockwell DJ, Zinober RC, Blake AW, Beddard GS, Olmsted PD, Radford SE Unfolding dynamics of proteins under applied force Philosophical Transactions of the Royal Society of London, Series A: Mathematical, Physical and Engineering Sciences 361 739-740, 2003
Zinober RC, Brockwell DJ, Beddard GS, Blake AW, Olmsted PD, Radford SE, Smith DA Mechanically unfolding proteins: The effect of unfolding history and the supramolecular scaffold PROTEIN SCI 11 2759-2765, 2002
Brockwell DJ, Beddard GS, Clarkson J, Zinober RC, Blake AW, Trinick J, Olmsted PD, Smith DAM, Radford SE The Effect of Core Destabilization on the Mechanical Resistance of I27 Biophysical Journal 83 458-472, 2002
Gell C, Brockwell DJ, Beddard GS, Radford SE, Kalverda AP, Smith DA Accurate use of single molecule fluorescence correlation spectroscopy to determine molecular diffusion times SINGLE MOL 2 177-181, 2001
Brockwell DJ, Smith DAM, Radford SE Protein folding mechanisms: new methods and emerging ideas Current Opinion in Structural Biology 16-25, 2000
Gell C, Brockwell D, Smith A Handbook of single molecule fluorescence spectroscopy,
Iadanza MG, Higgins AJ, Schiffrin R, Calabrese AN, Brockwell DJ, Ashcroft AE, Radford SE, Ranson NA Lateral opening in the intactβ-barrel assembly machinery captured by cryo-EM Nature Communications -,