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Professor Michael Geeves

Professor of Physical Biochemistry

School of Biosciences

 

Professor Mike Geeves joined the School of Biosciences in April 1999. He studied biochemistry as an undergraduate at the University of Birmingham then went on to the University of Bristol to work on a PhD with David Trentham. It was here that he first came to work on the myosin motor which has been the focus of his work ever since. In those early days it was muscle myosin - the only known form of myosin. After completing his PhD he spent 2 years at the University of California, Santa Cruz studying enzymology at sub-zero temperatures with Anthony Fink. He then return to spend 14 years at the University of Bristol working alongside Freddie Gutfreund, first as an SERC Junior Fellow then as a Royal Society University Fellow. At the end of the fellowship he moved to become a Group Leader in the new Max Planck Institute of Molecular Physiology that was being established in Dortmund by Roger Goody. He left there to take up the current position as Professor of Physical Biochemistry.

Mike is a member of the Mechanobiology group also known as MaDCaP.

ORCID ID: 0000-0002-9364-8898

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Article
Mijailovich, S. et al. (2017). Modeling the Actin.myosin ATPase cross-bridge cycle for skeletal and cardiac muscle myosin isoforms. Biophysical Journal [Online] 112:984-996. Available at: http://dx.doi.org/10.1016/j.bpj.2017.01.021.
Geeves, M. (2017). More Can Mean Less, or: Simplifying Sometimes Requires Ideas To Be More Complicated. Biophysical Journal [Online] 112:2467-2468. Available at: http://dx.doi.org/10.1016/j.bpj.2017.05.004.
Walklate, J. et al. (2016). The Most Prevalent Freeman-Sheldon Syndrome Mutations in the Embryonic Myosin Motor Share Functional Defects. Journal of Biological Chemistry [Online] 291:10318-10331. Available at: http://doi.org/10.1074/jbc.M115.707489.
Brooker, H., Geeves, M. and Mulvihill, D. (2016). Analysis of biophysical and functional consequences of Tropomyosin - fluorescent protein fusions. FEBS letters [Online]:3111-3121. Available at: http://onlinelibrary.wiley.com/doi/10.1002/1873-3468.12346/full.
Geeves, M. (2016). The ATPase mechanism of myosin and actomyosin. Biopolymers [Online] 105:483-491. Available at: http://doi.org/10.1002/bip.22853.
Walklate, J., Ujfalusi, Z. and Geeves, M. (2016). Myosin isoforms and the mechanochemical cross-bridge cycle. Journal of Experimental Biology [Online] 219:168-174. Available at: http://doi.org/10.1242/jeb.124594.
Mijailovich, S. et al. (2016). Three-dimensional stochastic model of actin–myosin binding in the sarcomere lattice. The Journal of General Physiology [Online] 148:459-488. Available at: http://doi.org/10.1085/jgp.201611608.
Geeves, M., Hitchcock-DeGregori, S. and Gunning, P. (2015). A systematic nomenclature for mammalian tropomyosin isoforms. Journal of Muscle Research and Cell Motility [Online] 36:147-153. Available at: http://doi.org/10.1007/s10974-014-9389-6.
Walklate, J. and Geeves, M. (2015). Temperature manifold for a stopped-flow machine to allow measurements from −10 to +40°C. Analytical Biochemistry [Online] 476:11-16. Available at: http://doi.org/10.1016/j.ab.2015.01.020.
Bloemink, M. et al. (2015). The Relay/Converter Interface Influences Hydrolysis of ATP by Skeletal Muscle Myosin II. Journal of Biological Chemistry [Online] 291:1763-1773. Available at: http://doi.org/10.1074/jbc.M115.688002.
Nag, S. et al. (2015). Contractility parameters of human -cardiac myosin with the hypertrophic cardiomyopathy mutation R403Q show loss of motor function. Science Advances [Online] 1:e1500511-e1500511. Available at: http://doi.org/10.1126/sciadv.1500511.
Lehman, W. et al. (2015). Phosphorylation of Ser283 enhances the stiffness of the tropomyosin head-to-tail overlap domain. Archives of Biochemistry and Biophysics [Online] 571:10-15. Available at: http://doi.org/10.1016/j.abb.2015.02.026.
Lehrer, S. and Geeves, M. (2014). The myosin-activated thin filament regulatory state, M − -open: a link to hypertrophic cardiomyopathy (HCM). Journal of Muscle Research and Cell Motility [Online] 35:153-160. Available at: http://doi.org/10.1007/s10974-014-9383-z.
Bloemink, M. et al. (2014). The Hypertrophic Cardiomyopathy Myosin Mutation R453C Alters ATP Binding and Hydrolysis of Human Cardiac beta-Myosin. Journal of Biological Chemistry [Online] 289:5158-5167. Available at: http://dx.doi.org/10.1074/jbc.M113.511204.
Lawrence, A. et al. (2014). FAD binding, cobinamide binding and active site communication in the corrin reductase (CobR). Bioscience Reports [Online] 34:345-355. Available at: http://dx.doi.org/10.1042/BSR20140060.
Desai, R., Geeves, M. and Kad, N. (2014). Using Fluorescent Myosin to Directly Visualize Cooperative Activation of Thin Filaments. Journal of Biological Chemistry [Online]:jbc.M114.609743-jbc.M114.609743. Available at: http://dx.doi.org/10.1074/jbc.M114.609743.
Geeves, M. and Lehrer, S. (2014). Cross-Talk, Cross-Bridges, and Calcium Activation of Cardiac Contraction. Biophysical Journal [Online] 107:543-545. Available at: http://dx.doi.org/10.1016/j.bpj.2014.06.019.
Janco, M. et al. (2013). Polymorphism in tropomyosin structure and function. Journal of Muscle Research and Cell Motility [Online] 34:177-187. Available at: http://dx.doi.org/10.1007/s10974-013-9353-x.
Bloemink, M. et al. (2013). The Superfast Human Extraocular Myosin Is Kinetically Distinct from the Fast Skeletal IIa, IIb, and IId Isoforms. Journal of Biological Chemistry [Online] 288:27469-27479. Available at: http://dx.doi.org/10.1074/jbc.M113.488130.
Li, X. et al. (2012). The flexibility of two tropomyosin mutants, D175N and E180G, that cause hypertrophic cardiomyopathy. Biochemical and Biophysical Research Communications [Online] 424:493-496. Available at: http://dx.doi.org/10.1016/j.bbrc.2012.06.141.
Janco, M. et al. (2012). α-Tropomyosin with a D175N or E180G Mutation in Only One Chain Differs from Tropomyosin with Mutations in Both Chains. Biochemistry [Online] 51:9880-9890. Available at: http://dx.doi.org/10.1021/bi301323n.
Deacon, J. et al. (2012). Identification of functional differences between recombinant human α and β cardiac myosin motors. Cellular and Molecular Life Sciences [Online] 69:2261-2277. Available at: http://dx.doi.org/10.1007/s00018-012-0927-3.
Canepari, M. et al. (2012). Actomyosin kinetics of pure fast and slow rat myosin isoforms studied by in vitro motility assay approach. Experimental Physiology [Online] 97:873-881. Available at: http://dx.doi.org/10.1113/expphysiol.2012.064576.
Kalyva, A., Schmidtmann, A. and Geeves, M. (2012). In Vitro Formation and Characterization of the Skeletal Muscle α·β Tropomyosin Heterodimers. Biochemistry [Online] 51:6388-6399. Available at: http://dx.doi.org/10.1021/bi300340r.
Geeves, M. (2012). 4.13 Thin Filament Regulation. Comprehensive Biophysics [Online] 4:251-267. Available at: http://dx.doi.org/10.1016/B978-0-12-374920-8.00416-1.
Mijailovich, S. et al. (2012). The Hill Model for Binding Myosin S1 to Regulated Actin Is not Equivalent to the McKillop–Geeves Model. Journal of Molecular Biology [Online] 417:112-128. Available at: http://dx.doi.org/10.1016/j.jmb.2012.01.011.
Deery, E. et al. (2012). An enzyme-trap approach allows isolation of intermediates in cobalamin biosynthesis. Nature Chemical Biology [Online] 8:933-940. Available at: http://dx.doi.org/10.1038/nchembio.1086.
Mijailovich, S. et al. (2012). Cooperative regulation of myosin-S1 binding to actin filaments by a continuous flexible Tm–Tn chain. European Biophysics Journal [Online] 41:1015-1032. Available at: http://dx.doi.org/10.1007/s00249-012-0859-8.
Deacon, J. et al. (2012). Erratum to: Identification of functional differences between recombinant human α and β cardiac myosin motors. Cellular and Molecular Life Sciences [Online] 69:4239-4255. Available at: http://dx.doi.org/10.1007/s00018-012-1111-5.
Geeves, M. and Ranatunga, K. (2012). Tuning the Calcium Sensitivity of Cardiac Muscle. Biophysical Journal [Online] 103:849-850. Available at: http://dx.doi.org/10.1016/j.bpj.2012.07.039.
Preller, M. et al. (2011). Structural Basis for the Allosteric Interference of Myosin Function by Reactive Thiol Region Mutations G680A and G680V. Journal of Biological Chemistry [Online] 286:35051-35060. Available at: http://dx.doi.org/10.1074/jbc.M111.265298.
Bloemink, M. et al. (2011). Two Drosophila myosin transducer mutants with distinct cardiomyopathies have divergent ADP and actin affinities. Journal of Biological Chemistry [Online] 286:28435-28443. Available at: http://dx.doi.org/10.1074/jbc.M111.258228.
Korte, F. et al. (2011). Upregulation of cardiomyocyte ribonucleotide reductase increases intracellular 2 deoxy-ATP, contractility, and relaxation. Journal of Molecular and Cellular Cardiology [Online] 51:894-901. Available at: http://dx.doi.org/10.1016/j.yjmcc.2011.08.026.
Bloemink, M. and Geeves, M. (2011). Shaking the myosin family tree: Biochemical kinetics defines four types of myosin motor. Seminars in Cell & Developmental Biology [Online] 22:961-967. Available at: http://dx.doi.org/10.1016/j.semcdb.2011.09.015.
Geeves, M. et al. (2011). Cooperative [Ca²+]-dependent regulation of the rate of myosin binding to actin: solution data and the tropomyosin chain model. Biophysical Journal [Online] 100:2679-2687. Available at: http://dx.doi.org/10.1016/j.bpj.2011.04.020.
Rayes, R. et al. (2011). Dynamics of tropomyosin in muscle fibers as monitored by saturation transfer EPR of bi-functional probe. PLoS ONE [Online] 6:e21277. Available at: http://dx.doi.org/10.1371/journal.pone.0021277.
Adamek, N., Geeves, M. and Coluccio, L. (2011). Myo1c mutations associated with hearing loss cause defects in the interaction with nucleotide and actin. Cellular and Molecular Life Sciences [Online] 68:139-150. Available at: http://dx.doi.org/10.1007/s00018-010-0448-x.
Adamek, N. et al. (2010). Modification of Loop 1 Affects the Nucleotide Binding Properties of Myo1c, the Adaptation Motor in the Inner Ear. Biochemistry [Online] 49:958-971. Available at: http://dx.doi.org/10.1021/bi901803j.
Johnson, M. et al. (2010). Targeted amino-terminal acetylation of recombinant proteins in E. coli. PLoS ONE [Online] 5:e15801. Available at: http://dx.plos.org/10.1371/journal.pone.0015801.
Mijailovich, S. et al. (2010). Resolution and uniqueness of estimated parameters of a model of thin filament regulation in solution. Computational Biology and Chemistry [Online] 34:19-33. Available at: http://dx.doi.org/10.1016/j.compbiolchem.2009.11.002.
Jenkins, D. et al. (2009). Rapid folding of the prion protein captured by pressure-jump. European Biophysics Journal [Online] 38:625-635. Available at: http://dx.doi.org/10.1007/s00249-009-0420-6.
Albet-Torres, N. et al. (2009). Drug Effect Unveils Inter-head Cooperativity and Strain-dependent ADP Release in Fast Skeletal Actomyosin. Journal of Biological Chemistry [Online] 284:22926-22937. Available at: http://dx.doi.org/10.1074/jbc.M109.019232.
Bloemink, M. et al. (2009). Alternative Exon 9-Encoded Relay Domains Affect More than One Communication Pathway in the Drosophila Myosin Head. Journal of Molecular Biology [Online] 389:707-721. Available at: http://dx.doi.org/10.1016/j.jmb.2009.04.036.
Bernstein, S. and Geeves, M. (2008). The XXXVII European muscle conference: Oxford September 2008. Journal of Muscle Research and Cell Motility [Online] 29:253-256. Available at: http://dx.doi.org/10.1007/s10974-008-9151-z.
Coulton, A. et al. (2008). Role of the Head-to-Tail Overlap Region in Smooth and Skeletal Muscle beta-Tropomyosin. Biochemistry [Online] 47:388-397. Available at: http://dx.doi.org/10.1021/bi701144g.
Meshcheryakov, V. et al. (2008). Crystallization and preliminary X-ray crystallographic analysis of full-length yeast tropomyosin 2 from Saccharomyces cerevisiae. Acta Crystallogr Section F-Structural Biology and Crystallization Communications [Online] 64:528-530. Available at: http://dx.doi.org/10.1107/S1744309108013110.
Adamek, N., Coluccio, L. and Geeves, M. (2008). Calcium sensitivity of the cross-bridge cycle of Myo1c, the adaptation motor in the inner ear. Proceedings of the National Academy of Sciences of the United States of America. [Online] 105:5710-5715. Available at: http://dx.doi.org/10.1073/pnas.0710520105 .
Smith, D. et al. (2008). Towards a Unified Theory of Muscle Contraction. I: Foundations. Annals of Biomedical Engineering [Online] 36:1624-1640. Available at: http://dx.doi.org/10.1007/s10439-008-9536-6.
Tsiavaliaris, G. et al. (2008). Mechanism, regulation, and functional properties of Dictyostelium myosin-1B. Journal of Biological Chemistry [Online] 283:4520-4527. Available at: http://dx.doi.org/10.1074/jbc.M708113200.
Maytum, R. et al. (2008). Ultra short yeast tropomyosins show novel Myosin regulation. Journal of Biological Chemistry [Online] 283:1902-1910. Available at: http://dx.doi.org/10.1074/jbc.M708593200.
Pearson, D., Swartz, D. and Geeves, M. (2008). Fast Pressure Jumps Can Perturb Calcium and Magnesium Binding to Troponin C F29W†. Biochemistry [Online] 47:12146-12158. Available at: http://dx.doi.org/10.1021/bi801150w.
Coulton, A. et al. (2007). Acetylation regulates tropomyosin function in the fission yeast Schizosaccharomyces pombe. Journal of Cell Science [Online] 120:1635-1645. Available at: http://dx.doi.org/10.1242/jcs.001115 .
Iorga, B., Adamek, N. and Geeves, M. (2007). The slow skeletal muscle isoform of myosin shows kinetic features common to smooth and non-muscle myosins. Journal of Biological Chemistry [Online] 282:3559-3570. Available at: http://www.jbc.org/cgi/reprint/282/6/3559.
Boussouf, S. et al. (2007). The regulation of Myosin binding to actin filaments by lethocerus troponin. Journal of Molecular Biology [Online] 373:587-98. Available at: http://dx.doi.org/10.1016/j.jmb.2007.07.066 .
Geeves, M. et al. (2007). A variable domain near the ATP-binding site in Drosophila muscle myosin is part of the communication pathway between the nucleotide and actin-binding sites. Journal of Molecular Biology [Online] 368:1051-1066. Available at: http://dx.doi.org/10.1016/j.jmb.2007.02.042.
Bloemink, M. et al. (2007). Kinetic Analysis of the Slow Skeletal Myosin MHC-1 Isoform from Bovine Masseter Muscle. Journal of Molecular Biology [Online] 373:1184-1197. Available at: http://dx.doi.org/10.1016/j.jmb.2007.08.050 .
Kintses, B. et al. (2007). Reversible movement of switch 1 loop of myosin determines actin interaction. EMBO Journal [Online] 26:265-274. Available at: http://www.nature.com/emboj/journal/v26/n1/pdf/7601482a.pdf.
Malnasi-Csizmadia, A. et al. (2007). Selective perturbation of the myosin recovery stroke by point mutations at the base of the lever arm affects ATP hydrolysis and phosphate release. Journal of Biological Chemistry [Online] 282:17658-17664. Available at: http://dx.doi.org/10.1074/jbc.M701447200.
Boussouf, S. et al. (2007). Role of tropomyosin isoforms in the calcium sensitivity of striated muscle thin filaments. Journal of Muscle Research and Cell Motility [Online] 28:49-58. Available at: http://www.springerlink.com/content/24jg471841626270/.
Coulton, A., Lehrer, S. and Geeves, M. (2006). Functional homodimers and heterodimers of recombinant smooth muscle tropomyosin. Biochemistry [Online] 45:12853-8. Available at: http://dx.doi.org/10.1021/bi0613224.
Rodger, A. et al. (2006). Looking at long molecules in solution: what happens when they are subjected to Couette flow? Physical Chemistry Chemical Physics [Online] 8:3161-71. Available at: http://dx.doi.org/10.1039/B604810M.
Adio, S. et al. (2006). Kinetic and mechanistic basis of the nonprocessive Kinesin-3 motor NcKin3. Journal of Biological Chemistry [Online] 281:37782-93. Available at: http://dx.doi.org/10.1074/jbc.M605061200.
Durrwang, U. et al. (2006). Dictyostelium myosin-IE is a fast molecular motor involved in phagocytosis. Journal of Cell Science [Online] 119:550-58. Available at: http://dx.doi.org/10.1242/​jcs.02774.
Nyitrai, M. et al. (2006). What Limits the Velocity of Fast-skeletal Muscle Contraction in Mammals? Journal of Molecular Biology [Online] 355:432-42. Available at: http://dx.doi.org/10.1016/j.jmb.2005.10.063.
Herm-Gotz, A. et al. (2006). Functional and biophysical analyses of the class XIV Toxoplasma gondii Myosin D. Journal of Muscle Research and Cell Motility 27:139-51.
Gewiss Mogensen, E. et al. (2006). Cryptococcus neoformans senses CO2 through the carbonic anhydrase Can2 and the adenylyl cyclase Cac1. Eukaryotic Cell [Online] 5:103-111. Available at: http://dx.doi.org/10.1128/EC.5.1.103-111.2006.
Kremneva, E. et al. (2006). Thermal unfolding of smooth muscle and nonmuscle tropomyosin alpha-homodimers with alternatively spliced exons. FEBS Journal [Online] 273:588-600. Available at: http://dx.doi.org/10.1111/j.1742-4658.2005.05092.x.
Nalavadi, V. et al. (2005). Kinetic Mechanism of Myosin IXB and the Contributions of Two Class IX-specific Regions. Journal of Biological Chemistry [Online] 280:38957-38968. Available at: http://www.jbc.org/cgi/content/abstract/280/47/38957.
Geeves, M. and Holmes, K. (2005). The molecular mechanism of muscle contraction. Fibrous Proteins: Muscle and Molecular Motors [Online] 71:161-193. Available at: http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B7CTK-4HC15BD-5&_user=125871&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000010239&_version=1&_urlVersion=0&_userid=125871&md5=461c86721444b98b516e70b4c96e6fcc.
Clark, R. et al. (2005). Loop 1 of transducer region in mammalian class I myosin, Myo1b, modulates actin affinity, ATPase activity, and nucleotide access. Journal of Biological Chemistry [Online] 280:30935-42. Available at: http://www.jbc.org/cgi/content/abstract/280/35/30935.
Fujita-Becker, S. et al. (2005). Changes in Mg2+ ion concentration and heavy chain phosphorylation regulate the motor activity of a class I myosin. Journal of Biological Chemistry [Online] 280:6064-71. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15579903.
Geeves, M., Fedorov, R. and Manstein, D. (2005). Molecular mechanism of actomyosin-based motility. Cellular and Molecular Life Sciences [Online] 62:1462-77. Available at: http://www.springerlink.com/content/j3q0w4gk15w58178/.
Nyitrai, M. and Geeves, M. (2004). Adenosine diphosphate and strain sensitivity in myosin motors. Philosophical Transactions of the Royal Society B: Biological Sciences [Online] 359:1867-77. Available at: http://dx.doi.org/10.1098%2Frstb.2004.1560.
Maytum, R. et al. (2004). Tropomyosin exon 6b is troponin-specific and required for correct acto-myosin regulation. Journal of Biological Chemistry [Online] 279:18203-9. Available at: http://dx.doi.org/10.1074/jbc.M311636200 .
Shimada, A. et al. (2004). The core FH2 domain of diaphanous-related formins is an elongated actin binding protein that inhibits polymerization. Molecular Cell [Online] 13:511-522. Available at: http://dx.doi.org/10.1016/S1097-2765(04)00059-0.
Kremneva, E. et al. (2004). Effects of two familial hypertrophic cardiomyopathy mutations in alpha-tropomyosin, Asp175Asn and Glu180Gly, on the thermal unfolding of actin-bound tropomyosin. Biophysical Journal [Online] 87:3922-33. Available at: http://dx.doi.org/10.1529/biophysj.104.048793 .
Attwood, P. and Geeves, M. (2004). Kinetics of an enzyme-catalyzed reaction measured by electrospray ionization mass spectrometry using a simple rapid mixing attachment. Analytical Biochemistry [Online] 334:382-9. Available at: http://dx.doi.org/10.1016/j.ab.2004.08.010 .
Crevel, I. et al. (2004). What kinesin does at roadblocks: the coordination mechanism for molecular walking. EMBO Journal [Online] 23:23-32. Available at: http://dx.doi.org/10.1038/sj.emboj.7600042.
Batters, C. et al. (2004). Myo1c is designed for the adaptation response in the inner ear. Embo Journal [Online] 23:1433-40. Available at: http://dx.doi.org/ 10.1038/sj.emboj.7600169 .
Silva, R., Sparrow, J. and Geeves, M. (2003). Isolation and kinetic characterisation of myosin and myosin S1 from the Drosophila indirect flight muscles. Journal of Muscle Research and Cell Motility [Online] 24:489-498. Available at: http://dx.doi.org/10.1023/B:JURE.0000009809.69829.74.
Smith, D., Maytum, R. and Geeves, M. (2003). Cooperative regulation of myosin-actin interactions by a continuous flexible chain I: actin-tropomyosin systems. Biophysical Journal [Online] 84:3155-3167. Available at: http://dx.doi.org/10.1016/S0006-3495(03)70040-X.
Miller, B. et al. (2003). Kinetic analysis of Drosophila muscle myosin isoforms suggests a novel mode of mechanochemical coupling. Journal of Biological Chemistry [Online] 278:50293-50300. Available at: http://dx.doi.org/10.1074/jbc.M308318200 .
Smith, D. and Geeves, M. (2003). Cooperative regulation of myosin-actin interactions by a continuous flexible chain II: actin-tropomyosin-troponin and regulation by calcium. Biophysical Journal [Online] 84:3168-3180. Available at: http://dx.doi.org/10.1016/S0006-3495(03)70041-1.
Maytum, R. et al. (2003). Differential regulation of the actomyosin interaction by skeletal and cardiac troponin isoforms. Journal of Biological Chemistry [Online] 278:6696-6701. Available at: http://dx.doi.org/10.1074/jbc.M210690200.
Clark, R. et al. (2003). Probing nucleotide dissociation from myosin in vitro using microgram quantities of myosin. Journal of Muscle Research and Cell Motility [Online] 24:315-321. Available at: http://dx.doi.org/10.1023/A:1025481908301.
Nyitrai, M., Szent-Gyorgyi, A. and Geeves, M. (2003). Interactions of the two heads of scallop (Argopecten irradians) heavy meromyosin with actin: influence of calcium and nucleotides. Biochemical Journal [Online] 370:839-848. Available at: http://dx.doi.org/10.1042/BJ20021519 .
Nyitrai, M. et al. (2003). Ionic interactions play a role in the regulatory mechanism of scallop heavy meromyosin. Biophysical Journal [Online] 85:1053-62. Available at: http://dx.doi.org/10.1016/S0006-3495(03)74544-5.
Nyitrai, M., Szent-Gyorgyi, A. and Geeves, M. (2002). A kinetic model of the co-operative binding of calcium and ADP to scallop (Argopecten irradians) heavy meromyosin. Biochemical Journal [Online] 365:19-30. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12071838 .
Geeves, M. and Lehrer, S. (2002). Modeling thin filament cooperativity. Biophysical Journal [Online] 82:1677-1679. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11898795 .
Maytum, R., Geeves, M. and Lehrer, S. (2002). A modulatory role for the troponin T tail domain in thin filament regulation. Journal of Biological Chemistry [Online] 277:29774-29780 . Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12045197 .
Pearson, D. et al. (2002). A novel pressure-jump apparatus for the microvolume analysis of protein-ligand and protein-protein interactions: its application to nucleotide binding to skeletal-muscle and smooth-muscle myosin subfragment-1. Biochemical Journal [Online] 366:643-651. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12010120 .
Tsiavaliaris, G. et al. (2002). Mutations in the relay loop region result in dominant-negative inhibition of myosin II function in Dictyostelium. EMBO Reports [Online] 3:1099-1105. Available at: http://dx.doi.org/10.1093/embo-reports/kvf214.
Geeves, M. (2002). Stretching the lever-arm theory. Nature [Online] 415:129-131. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11805818 .
Attwood, P. and Geeves, M. (2002). Changes in catalytic activity and association state of pyruvate carboxylase which are dependent on enzyme concentration. Archives of Biochemistry and Biophysics [Online] 401:63-72. Available at: http://dx.doi.org/10.1016/S0003-9861(02)00039-5.
Herm-Gotz, A. et al. (2002). Toxoplasma gondii myosin A and its light chain: a fast, single-headed, plus-end-directed motor. EMBO Journal [Online] 21:2149-2158. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11980712 .
Malnasi-Csizmadia, A. et al. (2001). Kinetic resolution of a conformational transition and the ATP hydrolysis step using relaxation methods with a Dictyostelium myosin II mutant containing a single tryptophan residue. Biochemistry [Online] 40:12727-12737. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11601998 .
Maytum, R. et al. (2001). Regulatory properties of tropomyosin effects of length, isoform, and N-terminal sequence. Biochemistry [Online] 40:7334-7341. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11401582 .
Lohmann, K. et al. (2001). Overexpression of human cardiac troponin in Escherichia coli: its purification and characterization. Protein Expression and Purification [Online] 21:49-59. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11162386 .
Weiss, S. et al. (2001). Differing ADP Release Rates from Myosin Heavy Chain Isoforms Define the Shortening Velocity of Skeletal Muscle Fibers. Journal of Biological Chemistry 276:45902-45908.
Berger, C. et al. (2001). ADP binding induces an asymmetry between the heads of unphosphorylated myosin. Journal of Biological Chemistry [Online] 276:23240-23245. Available at: http://dx.doi.org/10.1074/jcb.M100524200.
Rogers, K. et al. (2001). KIF1D is a fast non-processive kinesin that demonstrates novel K-loop-dependent mechanochemistry. Embo Journal [Online] 20:5101-5113. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11566875 .
Maytum, R., Geeves, M. and Lehrer, S. (2000). Effects of TnT and TnT1 upon regulation of actin-tropomyosin. Biophysical Journal 78:365A-365A.
Lehman, W. et al. (2000). Tropomyosin and actin isoforms modulate the localization of tropomyosin strands on actin filaments. Journal of Molecular Biology [Online] 302:593-606. Available at: http://dx.doi.org/10.1006/jmbi.2000.4080.
Weiss, S. et al. (2000). Kinetic characterisation of the myosin A from toxoplasma gondii. Biophysical Journal 78:1432Pos.
Coluccio, L. et al. (2000). Truncation of a mammalian myosin I results in loss of Ca2+-sensitive motility. Journal of Biological Chemistry [Online] 275:21618-21623. Available at: http://dx.doi.org/10.1074/jbc.M000363200.
Weiss, S., Chizhov, I. and Geeves, M. (2000). A flash photolysis fluorescence/light scattering apparatus for use with sub microgram quantities of muscle proteins. Journal of Muscle Research and Cell Motility [Online] 21:423-432. Available at: http://dx.doi.org/10.1023/A:1005690106951.
Geeves, M., Perreault-Micale, C. and Coluccio, L. (2000). Kinetic analyses of a truncated mammalian myosin I suggest a novel isomerization event preceding nucleotide binding. Journal of Biological Chemistry [Online] 275:21624-21630. Available at: http://dx.doi.org/10.1074/jbc.M000342200.
Maytum, R., Geeves, M. and Konrad, M. (2000). Actomyosin regulatory properties of yeast tropomyosin are dependent upon N-terminal modification. Biochemistry [Online] 39:11913-11920. Available at: http://dx.doi.org/10.1021/bi000977g.
Furch, M. et al. (2000). Stabilization of the actomyosin complex by negative charges on myosin. Biochemistry [Online] 39:11602-11608. Available at: http://dx.doi.org/10.1021/bi000985x.
Holmes, K. and Geeves, M. (2000). The structural basis of muscle contraction. Philosophical Transactions of the Royal Society B: Biological Sciences [Online] 355:419-431. Available at: http://dx.doi.org/10.1098/rstb.2000.0583.
Furch, M., Geeves, M. and Manstein, D. (2000). Kinetic dissection of the effect of negative charges of loop 2 on the enzymatic activity of myosin. Biophysical Journal 78:393-393.
Geeves, M., Chai, M. and Lehrer, S. (2000). Inhibition of actin-myosin subfragment 1 ATPase activity by troponin I and IC: Relationship to the thin filament states of muscle. Biochemistry [Online] 39:9345-9350. Available at: http://dx.doi.org/10.1021/bi0002232.
Book section
Adamek, N. and Geeves, M. (2014). Use of pyrene-labelled actin to probe actin-Myosin interactions: kinetic and equilibrium studies. in: Fluorescent Methods Applied to Molecular Motors: from single molecules to whole cells. Springer, pp. 87-104. Available at: http://link.springer.com/book/10.1007%2F978-3-0348-0856-9.
Toseland, C. and Geeves, M. (2014). Rapid reaction kinetic techniques. in: Toseland, C. P. and Fili, N. eds. Fluorescent Methods Applied to Molecular Motors: from single molecules to whole cells. Springer, pp. 49-64. Available at: http://dx.doi.org/10.1007/978-3-0348-0856-9.
Geeves, M. and Pearson, D. (2013). Kinetics: Relaxation Methods. in: Roberts, G. C. K. ed. Encyclopedia of Biophysics . Springer Berlin Heidelberg, pp. 1207-1212. Available at: http://dx.doi.org/10.1007/978-3-642-16712-6_62.
Johnson, M., Geeves, M. and Mulvihill, D. (2013). Production of Amino-Terminally Acetylated Recombinant Proteins in E. coli. in: Hake, S. and Janzen, C. eds. Protein Acetylation. Humana Press, pp. 193-200. Available at: http://dx.doi.org/10.1007/978-1-62703-305-3_15.
Geeves, M. and Lehrer, S. (2002). Cooperativity in the Ca2+ regulation of muscle contraction. in: Molecular Interactions of Actin. Springer-Verlag Berlin and Heidelberg GmbH & Co. K , pp. 111-132. Available at: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11892276 .
Conference or workshop item
Boussouf, S. and Geeves, M. (2007). Tropomyosin and troponin cooperativity on the thin filament. in: 33rd Symposium on Regulatory Mechanisms of Striated Muscle Contraction . Berlin, Germany: Springer-Verlag Berlin, Heidelberger Platz 3, D-14197 Berlin, Germany, pp. 99-109. Available at: http://dx.doi.org/10.1007/978-4-431-38453-3_10.
Showing 118 of 121 total publications in KAR. [See all in KAR]

 

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Molecular Motors

ccycle movie

Myosins comprise one of the three major families of molecular motors which operate in cells. In addition to muscle contraction they are involved in a wide range cellular processes such as cell division, phagacytosis and vesicle transport. The understanding of the molecular events which make up the ATP driven cross bridge cycle (see fig) advances year by year yet there are continually new surprises in the variety and range of activities shown by members of the super family. We know know that myosins can move in opposite directions along actin filaments, can be processive and can act as cytoskeletal cross-linkers and strain sensors. Despite this variety in behaviour the same basic myosin motor domain brings about the different activities. Our work is directed towards understanding how the mechanochemistry of the myosin motor domain is tuned to produce these widely differing activities and how the motor activity is regulated.

 

 

Current Projects:

Mechanochemical coupling in the Myosin super family

 

The cross bridge cycle can be described as a series of coupled biochemical and mechanical events. The efficient conversion of biochemical energy requires a very precise temporal coupling between the biochemical and mechanical events and the structure of the myosin head is presumably designed to achieve this end. The cross bridge cycle for fast skeletal muscle is the most thoroughly studied system and the description of the cycle that follows refers to this actomyosin. The basic cycle is believed to be the same for all myosins but the relative rates of individual steps are altered by changes in the amino acid sequence of myosin to tune each myosin for its particular physiological role. Comparisons of the properties of different myosins together with mutagenesis of specific amino acid residues or of short sequences are the most active areas of current research.

xbcycle.jpeg

Colour code for the Figure: Actin monomers - orange; Myosin head - green; Light chains - yellow; Myosin S2 & thick filament - black; ATP/ADP - blue/pink; Pi - pink/yellow.

The essential mechanochemical steps are shown lettered (a) to (e). ATP binding to either a rest length myosin head (c) or to a head bearing a load (b) results in a change in conformation of the myosin head causing a rapid almost irreversible dissociation of the myosin head from actin (d). Following detachment from actin the ATP is hydrolysed to ADP and Pi, both of which remain very tightly bound to the myosin head (e). The hydrolysis is relatively rapid (taking about 10 msec) and reversible (Keq = 10). The small value of the Keq indicates that the free energy of ATP hydrolysis is not released but remains within the structure of the M.ADP.Pi complex. The crystal structures of myosin heads with nucleotides or nucleotide analogues bound are believed to represent one or other of these two structures (d & e). This suggests that the hydrolysis is accompanied by a major conformational change which represents the reversal or a re-priming of the power stroke. Structures d & e are quite stable and ADP and Pi will remain bound to the myosin head until the myosin binds to an actin site. The affinity of M.ADP.Pi for actin is significantly higher than that of M.ATP and if an actin site is within reach of the myosin head it will bind rapidly and reversibly to the actin site and in doing so can explore several potential actin binding sites. Current opinion suggests that the majority (>80%) of myosin heads within an active isometric (not shortening) muscle are in equilibrium between states a d & e.

When the myosin head binds to actin the interaction with actin can promote a major change in conformation (the power stroke) which is accompanied by the dissociation of Pi. Crystal structures suggest that the power stroke consists of a reorientation of part of the myosin head distal to the actin-binding site and includes the converter domain and the light chain-binding region (LCBD). This results in the displacement of the distal tip of the LCBD by up to 10 nm. The structural changes in the actin-myosin interface that produces the power stroke remain undefined.

If the filaments carry an external load then the power stroke results in the distortion of an elastic element (b. The location and nature of the elastic element is unknown but may represent a distortion in the myosin head or of the LCBD. For simplicity the elastic element is drawn here as part of the connection between the myosin head and the thick filament. While the myosin head carries a load and is elastically distorted the dissociation of Pi is a reversible event and Pi can rebind to reverse the power stroke (and also back through intermediates e & d).
If the external load is small then the power stroke results in the relative sliding of the actin and myosin filaments by a distance of up to 10 nm. Following the sliding ADP is released very quickly (within 2 msec) to be replaced within a msec by ATP and the myosin head dissociates once more to complete the cycle. The final element of the mechanochemical coupling is believed to be a mechanism to limit the rate of release of ADP until the sliding motion is complete. Thus ADP release from b is much slower than from c. The mechanism of the strain limited ADP release is under current investigation but appears to be a key event which differs between myosins designed for efficient fast shortening vs. efficient load bearing. The structural changes observed on binding ADP to complexes of actin with smooth muscle myosin and brush border myosin I might reflect this strain sensing mechanism.

The mechanical model of the cycle has been updated more recently as more detailed structural information has become available.

Regulation of myosin motors

The regulation of the interaction between actin (A) and myosin heads (M ) in skeletal muscle is brought about by a Ca2+ induced change in state of the thin filament proteins tropomyosin (Tm) and troponin (Tn).1,2. The thin filament is a repeating structure made up of units of Actin7.TmTn. Skeletal tropomyosin is a linear coiled coil structure approximately 400 Å long which interacts with 7 actin subunits and the troponin regulatory complex. This structural repeat containing 7 actins per unit was used by Hill et. al.3 as the cooperative unit size in their model of thin filament cooperativity.

McKillop & Geeves4 more recently proposed that the regulation of TmTn containing thin filaments can be interpreted in terms of a rapid Ca2+ dependent equilibrium between 3 states of the thin filament, blocked, closed and open. A steric blocking model of thin filament regulation has been proposed (Figure) which shows the relationship between the three states of the thin filament and the two step binding of S1 to actin.5, 6, 7

3state.jpg

1. Lehrer, S.S. (1994) J. Musc. Res. Cell Motil. 15, 232-236

2. Lehrer, S.S. and Geeves, M.A. (1998) J. Mol Biol. 277, 1081-1089

3. Hill, T.L., Eisenberg, E., and Greene, L. (1980) Proc. Natl. Acad. Sci USA 77, 3186-3190.

4. McKillop, D.F.A. and Geeves, M.A. (1991) Biochem. J. 279, 711-718.

5. Geeves, M.A., Goody, R.S. and Gutfreund H. (1984) J. Musc. Res. Cell. Motil. 5, 351-361.

6. Geeves, M.A. (1991) Biochem. J. 274, 1-14.

7. Geeves and Connibear (1995) Biophys. J. 68(4 Suppl), 194S-199S.

Inherited diseases of cardiac and skeletal muscle

There are a series of inherited human diseases of muscle caused by mutations in sarcomeric proteins such as the cardiomyopathies (hypertrophic and dilated cardiomyopathies) which can be life threatening or Freeman-Sheldon Syndrome which can cause developmental problems and major musculoskeletal deformities. The majority the cardiac mutations are found in myosin (~40% of known mutations) or myosin binding protein C (40%) with the rest in actin, tropomyosin, troponin, titin and other minor proteins. In most cases how the mutations cause the disease is not known.

Isolating or making the normal protein and the protein carrying the damaging mutation is one approach to understanding how the disease develops. Studying the protein in isolation can give some insight in to the problems caused by the mutation but studying the same protein in the context of its interacting partners provides much more detail whether this is in complex mixtures of proteins or in a contracting muscle.

We are studying the role of disease causing mutations in tropomyosin, troponin C, myosin and myosin binding protein C. Using the methods outlined elsewhere we can isolate the protein and reassemble the protein in complexes from filaments, myofibrils or in transgenic model systems like Drosophila flight muscle.

Development of novel fast reaction methods in skeletal and cardiac muscle contractiion

Pressure Jump

Rapid changes in pressure can be used to perturb the equilibrium position of many protein-protein or protein-ligand interactions on a time scale from 100 µs to several hundred seconds. Older equipment used volumes (1-2 mL) which are too extravagant for many biological materials. We have built a micro-volume pressure jump apparatus [Pearson et al. (2002) Biochem. J. 366(2): 643-651] based on the design of Clegg & Maxwell (1976) Rev. Sci. Inst. 47(11): 1383-93. The sample volume of the apparatus is 50 µL and it requires a minimum of 80 µL of sample for convenience of handling. The apparatus is designed with high pressure taps which allow simple rapid exchange of samples. Pressure is applied by means of a piezoelectric crystal stack separated from the sample by a polyimide membrane. Pressure jumps of up to +400 bar can be applied within 100 µs. The use of a piezoelectric stack allows the results of both pressure application and release to be repeatedly recorded and averaged, which allows small signal changes to be measured.

fast p jump

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Fast Pressure Jump Equipment. A piston, driven by a piezoelectric stack, allows fast repetitive pressure jumps to be made on 50 µL of sample solution held within a sapphire ring. Typically, a photomultiplier measures fluorescence at 90° to the incident light (a simple rearrangement allows measurement of transmitted light instead).

Flash photolysis

Transient kinetic methods such as stopped flow and quenched flow have been used to elucidate many of the fundamental features of the molecular interactions which underlie muscle contraction. However, these methods traditionally require relatively large amounts of protein (10-3g) and so have been used most effectively for the proteins purified from bulk muscle tissue of large animals or where the proteins can be expressed in large amounts (e.g., Dictyostelium). Single molecule and in vitro motility methods can be used on much smaller quantities but currently the range of measurements that can be made is limited. We have been developing methods which allow such fast transients to be studied on the much smaller quantities of protein (10-7, 10-6g) available from individual Drosophila muscles, single mammalian muscle fibers, human cardiac biopsies and non-muscle myosins.

Put the content of the callout in here. Flash Photolysis set up

 

 

 

 

 

 

 

 

 

 

 

 

 


Flash Photolysis set-up: Simultaneous time resolved detection of transmission changes and either fluorescence or light scattering changes, for which different light sources and a different set of filters are used.

The quartz cuvette in the core of the system operates with a minimal sample volume of 10 µl. A Laser System with a wavelength of 347 nm provides the source of the flashlight causing the fast release of ATP from a caged compound into the solution. The optical bench is designed for simultaneous time resolved detection of the following transmission changes and of fluorescence (or light scattering) changes (90°) in the cuvette to monitor the reactions kinetics. For detection of fluorescence changes a Xe-Hg-lamp is used, while for detection of light scattering a halogen lamp is prefered. The detection electronics include two photomultipliers and a digital oscilloscope connected to a PC.

Acknowledgements:

Major Current Collaborations:

  • Leslie Leinwand (University of Colorado, USA [http://biofrontiers.colorado.edu/about/directory/leinwand]),
  • Mike Regnier (University of Washington, USA [http://www.bioeng.washington.edu/regnier/people/regnier.html]),
  • Srboljub Mijailovich & Tom Irving (Illinois Institute of Technology, USA [https://science.iit.edu/people/faculty/thomas-irving]).
  • Corrado Poggesi [http://www.unifi.it/p-doc2-2013-200009-P-3f2a3c2d332e2c-0.html] & Chira Tesi (University of Florence, Italy [http://www.unifi.it/p-doc2-2013-200009-T-3f2a3d2c342b2e-0.html])

Previous long-term collaborators:

  • Drs Sam Lehrer & Lynne Coluccio, Boston Biomedical Research Institute
  • Deitmar Manstein, Hannover Medical School [https://www.mh-hannover.de/bpc_prof_manstein.html]
  • Rob Cross, University of Warwick [http://www2.warwick.ac.uk/fac/med/staff/cross/],
  • David Smith Monash University Melbourne, and Sanford Bernstein, San Diego State University [http://bernstein-lab.sdsu.edu/index.html]
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Year 1 BI300 Introduction to Biochemistry (Module convenor)

Year 2 BI503 Cell Biology

Final Year BI674 Molecular Machines in Biology (Module convenor)

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Member of editorial board for:

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PhD students

Cassidy Mackenzie

Chloe Johnson

 

 

 

 

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Enquiries: Phone: +44 (0)1227 823743

School of Biosciences, University of Kent, Canterbury, Kent, CT2 7NJ

Last Updated: 18/05/2017