Faculty of Biological Sciences

Dr Michael Harrison

BSc, PhD Oct 1990, Leeds
Lecturer
School of Biomedical Sciences

Background: Postdoctoral work: University of California-Berkeley, 1990-91; University of Leeds, 1991-1994. Wellcome Trust Career Development Fellowship, 1994-1998.

Contact:  Garstang 6.44a, +44(0) 113 34 37766, email address for  

Research Interests

Structure, function and molecular mechanisms of membrane protein complexes

Prostate cancer cell

The vacuolar H+-ATPase (V-ATPase) is a membrane protein complex that works as an ATP-fuelled acid pump. It plays a central role in cell physiology, with defects in it causing bone and kidney diseases, and compounds that affect it having potential as anti-cancer drugs. The V-ATPase is a molecular-scale machine: Two rotary motors (one consuming ATP 'fuel', the other pumping acid) are linked by a rotating axle that drives the acid pump in the cell membrane. The nature of this mechanical coupling and how it actually pumps the acid remain unanswered but fundamental questions. Tackling them is a key aim of the research in my lab, but this needs higher resolution structural models of the complex. To do this we work with structural biologists and use high-resolution electron microscopy to look at the 3-D organisation of the protein (see Muench et al (2011) Q. Rev. Biophys. 44, 311; Rawson et al (2015) Structure 23, 461).

 

The V-ATPase also connects to, and is controlled by, other cellular energy pathways. How it makes the connection, and how this controls the V-ATPase, are uncertain but appear to involve protein phosphorylation signals. We are also looking at the connection between cell signalling and ATPase activity, working out in particular the changes at the molecular level that cause the ATPase to become switched off (see Muench et al (2014) J. Mol. Biol. 426, 286).

In cultured cancer cells, some forms of the V-ATPase are present in large numbers at the cell surface where they appear to play a role in helping the cell maintain a stable internal pH. We are examining how this plasma membrane localisation comes about, and the cause-and-effect relationship it might have with tumour invasiveness (see Smith et al (2016) J. Biol. Chem. 291, 8500).

Receptor tyrosine kinases are a class of membrane proteins that play crucial roles in transmitting signals from growth factors and hormones into the cell, bringing about major changes in features such cell physiology, motility, growth and regeneration. Whilst we know a great deal about the detailed structures of parts of these complex molecules from x-ray crystallography, we currently don't have detailed structures for the whole receptor. Bringing together specialists in RTK cell biology and biochemistry with experts in cryo-electron microscopy, our objective is to solve the complete structures of important RTKs in both the silent and fully active states. The aim is to achieve a better understanding of the steps at the molecular level that link growth factor/hormone binding at the cell surface to the initiation of signalling within that cell.

 

Current Projects

1. Structure, function and regulation of the V-ATPase

2. Control of V-ATPase function by protein kinase signalling.

3. Role of the V-ATPase in cancer: isoform function, location and navigation around the cell

4. The role of ubiquitination in Vascular Endothelial Growth Factor Receptor -1/2 function (with Dr Vas Ponnambalam).

5. Structure of Receptor Tyrosine Kinases by high-resolution cryo-electron microscopy (with Dr Stephen Muench & Dr Vas Ponnambalam).

 

Faculty Research and Innovation



Studentship information

Postgraduate studentship areas:

  • High resolution cryo-electron microscopy of receptor tyrosine kinases (with Dr Stephen Muench and Dr Vas Ponnambalam)
  • All hands to the pump: what controls V-ATPase isoform activity at the plasma membrane of invasive cancer cells?
  • Regulation of the human v-ATPase pump by protein kinase activity

See also:

Modules managed

BMSC1103 - Basic Laboratory and Scientific Skills
BMSC2233 - Topics in Medical Sciences
BMSC3146 - Advanced Topics in Medical Sciences I
BMSC3236 - Advanced Topics in Medical Sciences II

Modules taught

BIOL2111/BIOC2301 - Integrated Biochemistry/Genetic Engineering
BIOL5372M - Advanced Biomolecular Technologies
BMSC1103 - Basic Laboratory and Scientific Skills
BMSC1110/SPSC1220 - Foundation modules
BMSC1213 - Basic Laboratory and Scientific Skills 2
BMSC2233 - Topics in Medical Sciences
BMSC3301 - Research Project in Biomedical Sciences
BMSC3399 - Extended Research Project Preparation
BMSC5382M - Extended Research Project
MEDI1216 - Introduction to Medical Sciences

Committees

Member of Undergraduate School Taught Student Education Committee (Programme Manager: Medical Sciences)

Group Leader Dr Michael Harrison  (Lecturer)

Structure, function and molecular mechanisms of membrane protein complexes 

Postgraduates

Joanna Mitchell (Co-supervisor) 20% FTE
Barnaby Roper (Co-supervisor) 33% FTE
Faheem Shaik (Co-supervisor) 35% FTE

Smith GA, Howell GJ, Phillips C, Muench SP, Ponnambalam S, Harrison MA Extracellular and luminal pH regulation by vacuolar H<sup>+</sup>-ATPase isoform expression and targeting to the plasma membrane and endosomes Journal of Biological Chemistry 291 8500-8515, 2016
DOI:10.1074/jbc.M116.723395
View abstract

Rawson S, Phillips C, Huss M, Tiburcy F, Wieczorek H, Trinick J, Harrison MA, Muench SP Structure of the vacuolar H+-ATPase rotary motor reveals new mechanistic insights Structure 23 461-471, 2015
DOI:10.1016/j.str.2014.12.016
View abstract

Muench SP, Scheres SHW, Huss M, Phillips C, Vitavska O, Wieczorek H, Trinick J, Harrison MA Subunit positioning and stator filament stiffness in regulation and power transmission in the V1 motor of the Manduca sexta V-ATPase Journal of Molecular Biology 426 286-300, 2014
DOI:10.1016/j.jmb.2013.09.018
View abstract

Muench SP, Trinick J, Harrison MA Structural divergence of the rotary ATPases Quarterly Reviews of Biophysics 44 311-356, 2011
DOI:10.1017/S0033583510000338
View abstract

Muench SP, Huss M, Song CF, Phillips C, Wieczorek H, Trinick J, Harrison MA Cryo-electron Microscopy of the Vacuolar ATPase Motor Reveals its Mechanical and Regulatory Complexity J MOL BIOL 386 989-999, 2009
DOI:10.1016/j.jmb.2009.01.014

Smith GA, Fearnley GW, Abdul-Zani I, Wheatcroft SB, Tomlinson DC, Harrison MA, Ponnambalam S Ubiquitination of basal VEGFR2 regulates signal transduction and endothelial function. Biol Open -, 2017
DOI:10.1242/bio.027896
View abstract

Tiede C, Bedford R, Heseltine SJ, Smith G, Wijetunga I, Ross R, Alqallaf D, Roberts APE, Balls A, Curd A, Hughes RE, Martin H, Needham SR, Zanetti-Domingues LC, Sadigh Y, Peacock TP, Tang AA, Gibson N, Kyle H, Platt GW, Ingram N, Taylor T, Coletta LP, Manfield I, Knowles M, Bell S, Esteves F, Maqbool A, Prasad RK, Drinkhill M, Bon RS, Patel V, Goodchild SA, Martin-Fernandez M, Owens RJ, Nettleship JE, Webb ME, Harrison M, Lippiat JD, Ponnambalam S, Peckham M, Smith A, Ferrigno PK, Johnson M, McPherson MJ, Tomlinson DC Affimer proteins are versatile and renewable affinity reagents eLife 6 -, 2017
DOI:10.7554/eLife.24903
View abstract

Rawson S, Harrison MA, Muench SP Rotating with the brakes on and other unresolved features of the vacuolar ATPase Biochemical Society Transactions 44 851-855, 2016
DOI:10.1042/BST20160043
View abstract

Fearnley GW, Smith GA, Abdul-Zani I, Yuldasheva N, Mughal NA, Homer-Vanniasinkam S, Kearney MT, Zachary IC, Tomlinson DC, Harrison MA, Wheatcroft SB, Ponnambalam S VEGF-A isoforms program differential VEGFR2 signal transduction, trafficking and proteolysis Biology Open 5 571-583, 2016
DOI:10.1242/bio.017434
View abstract

Smith GA, Fearnley GW, Abdul-Zani I, Wheatcroft SB, Tomlinson DC, Harrison MA, Ponnambalam S VEGFR2 Trafficking, Signaling and Proteolysis is Regulated by the Ubiquitin Isopeptidase USP8 Traffic 17 53-65, 2016
DOI:10.1111/tra.12341
View abstract

Smith GA, Tomlinson DC, Harrison MA, Ponnambalam S Ubiquitin-Mediated Regulation of Cellular Responses to Vascular Endothelial Growth Factors Progress in Molecular Biology and Translational Science -, 2016
DOI:10.1016/bs.pmbts.2016.02.006
View abstract

Smith GA, Fearnley GW, Tomlinson DC, Harrison MA, Ponnambalam S The cellular response to vascular endothelial growth factors requires co-ordinated signal transduction, trafficking and proteolysis Bioscience Reports 35 -, 2015
DOI:10.1042/BSR20150171
View abstract

Smith GA, Fearnley GW, Harrison MA, Tomlinson DC, Wheatcroft SB, Ponnambalam S Vascular endothelial growth factors: multitasking functionality in metabolism, health and disease. Journal of Inherited Metabolic Disease 38 753-763, 2015
DOI:10.1007/s10545-015-9838-4
View abstract

Karpov OA, Fearnley GW, Smith GA, Kankanala J, McPherson MJ, Tomlinson DC, Harrison MA, Ponnambalam S Receptor tyrosine kinase structure and function in health and disease AIMS Biophysics 2 476-502, 2015
DOI:10.3934/biophy.2015.4.476
View abstract

Richardson RA, Papachristos K, Read DJ, Harlen OG, Harrison M, Paci E, Muench SP, Harris SA Understanding the apparent stator-rotor connections in the rotary ATPase family using coarse-grained computer modeling Proteins: Structure, Function, and Bioinformatics 82 3298-3311, 2014
DOI:10.1002/prot.24680
View abstract

Muench SP, Rawson S, Eyraud V, Delmas AF, Da Silva P, Phillips C, Trinick J, Harrison MA, Gressent F, Huss M PA1b inhibitor binding to subunits C and E of the vacuolar ATPase reveals its insecticidal mechanism Journal of Biological Chemistry 289 16399-16408, 2014
DOI:10.1074/jbc.M113.541250
View abstract

Fearnley GW, Smith GA, Odell AF, Latham AM, Wheatcroft SB, Harrison MA, Tomlinson DC, Ponnambalam S Vascular endothelial growth factor A-stimulated signaling from endosomes in primary endothelial cells. Methods Enzymol 535 265-292, 2014
DOI:10.1016/B978-0-12-397925-4.00016-X
View abstract

Song CF, Papachristos K, Rawson S, Huss M, Wieczorek H, Paci E, Trinick J, Harrison MA, Muench SP Flexibility within the rotor and stators of the vacuolar H<sup>+</sup>-ATPase PLoS ONE 8 -, 2013
DOI:10.1371/journal.pone.0082207
View abstract

Fearnley GW, Smith GA, Harrison MA, Wheatcroft SB, Tomlinson DC, Ponnambalam S The biochemistry of new blood vessel formation in health and disease OA Biochemistry 1 -, 2013

Kuruvatti J, Prasad V, Williams R, Harrison MA, Jones RPO Motivations for donating blood and reasons why people lapse or never donate in Leeds, England: A 2001 questionnaire-based survey Vox Sanguinis 101 333-338, 2011
DOI:10.1111/j.1423-0410.2011.01488.x
View abstract

Jones RPO, Durose LJ, Phillips C, Keen JN, Findlay JBC, Harrison MA A site-directed cross-linking approach to the characterization of subunit E-subunit G contacts in the vacuolar H+-ATPase stator MOL MEMBR BIOL 27 147-159, 2010
DOI:10.3109/09687681003796441

Kóta Z, Páli T, Dixon N, Kee TP, Harrison MA, Findlay JBC, Finbow ME, Marsh D Incorporation of transmembrane peptides from the vacuolar H<sup>+</sup>-ATPase in phospholipid membranes: Spin-label electron paramagnetic resonance and polarized infrared spectroscopy Biochemistry 47 3937-3949, 2008
DOI:10.1021/bi7025112
View abstract

Dixon N, Pali T, Kee TP, Ball S, Harrison MA, Findlay JBC, Nyman J, Vaananen K, Finbow ME, Marsh D Interaction of spin-labeled inhibitors of the vacuolar H+-ATPase with the transmembrane V-o-sector BIOPHYS J 94 506-514, 2008
DOI:10.1529/biophysj.107.111781

Dixon N, Pali T, Kee TP, Ball S, Harrison MA, Findlay JBC, Nyman J, Vaananen K, Finbow ME, Marsh D Interaction of spin-labeled inhibitors of the vacuolar H+-ATPase with the transmembrane V-o-sector BIOPHYSICAL JOURNAL 94 506-514, 2008
DOI:10.1529/biophysj.107.111781

Dixon N, Pali T, Kee T, Ball SK, Harrison MA, Findlay JBC, Nyman J, Vaananen K, Finbow ME, Marsh D Interaction of spin-labelled inhibitors of the vacuolar H+-ATPase with the transmembrane Vo-sector Biophysical Journal 94 506-514, 2008
DOI:10.1529/biophysj.107.111781

Duarte AMS, Wolfs CJAM, van Nuland NAJ, Harrison MA, Findlay JBC, van Mierlo CPM, Hemminga MA Structure and localization of an essential transmembrane segment of the proton translocation channel of yeast H+-VATPase BBA-BIOMEMBRANES 1768 218-227, 2007
DOI:10.1016/j.bbamem.2006.07.014

Clare DK, Orlova EV, Finbow MA, Harrison MA, Findlay JBC, Saibil HR An expanded and flexible form of the vacuolar ATPase membrane sector STRUCTURE 14 1149-1156, 2006
DOI:10.1016/j.str.2006.05.014

Whyteside G, Meek PJ, Ball SK, Dixon N, Finbow ME, Kee TP, Findlay JBC, Harrison MA Concanamycin and indolyl pentadieneamide inhibitors of the vacuolar H+-ATPase bind with high affinity to the purified proteolipid subunit of the membrane domain BIOCHEMISTRY-US 44 15024-15031, 2005
DOI:10.1021/bi051529h

Jones RPO, Durose LJ, Findlay JBC, Harrison MA Defined sites of interaction between subunits E (Vma4p), C (Vma5p), and G (Vma10p) within the stator structure of the vacuolar H+-ATPase (vol 44, pg 3933, 2005) BIOCHEMISTRY-US 44 11924-11924, 2005
DOI:10.1021/bi058022r

Jones RPO, Durose LJ, Findlay JBC, Harrison MA Defined sites of interaction between subunits E (Vma4p), C (Vma5p), and G (Vma10p) within the stator structure of the vacuolar H+-ATPase (vol 44, pg 3933, 2005) BIOCHEMISTRY 44 11924-11924, 2005
DOI:10.1021/bi058022r

Jones RPO, Durose LJ, Findlay JBC, Harrison MA Erratum: Defined sites of interaction between subunits E (Vma4p), C (Vma5p), and G (Vma10p) within the stator structure of the vacuolar H +-ATPase (Biochemistry (March 15, 2005) 44, 10 (3933-3941)) Biochemistry 44 11924-, 2005
DOI:10.1021/bi058022r

Jones RPO, Durose LJ, Findlay JBC, Harrison MA Defined sites of interaction between subunits E (Vma4p), C (Vma5p), and G (Vma10p) within the stator structure of the vacuolar H+-ATPase BIOCHEMISTRY-US 44 3933-3941, 2005
DOI:10.1021/bi048402x

Pali T, Whyteside G, Dixon N, Kee TP, Ball S, Harrison MA, Findlay JBC, Finbow ME, Marsh D Interaction of inhibitors of the vacuolar H+-ATPase with the transmembrane V-o-sector BIOCHEMISTRY-US 43 12297-12305, 2004
DOI:10.1021/bi0493867

Harrison MA, Durose L, Song CF, Barratt E, Trinick J, Jones R, Findlay JBC Structure and function of the vacuolar H+-ATPase: moving from low-resolution models to high-resolution structures. Journal of Bioenergetics and Biomembranes 35 337-345, 2003
DOI:10.1023/A:1025728915565

Dixon N, Pali T, Ball S, Harrison MA, Marsh D, Findlay JBC, Kee TP New biophysical probes for structure-activity analyses of vacuolar-H+-ATPase enzymes ORG BIOMOL CHEM 1 4361-4363, 2003
DOI:10.1039/b311401e

Jones RPO, Prasad V, Kuruvatti J, Tahir N, Whitaker P, Dawson ASJ, Harrison MA, Williams R Remuneration for blood donation and attitudes towards blood donation and receipt in Leeds. Transfusion medicine 13 131-140, 2003
DOI:10.1046/j.1365-3148.2003.00433.x

Findlay JBC, Harrison MA A protein chemical approach to channel structure and function: the proton channel of the vacuolar H+-ATPase Ion Channels: from Atomic Resolution Physiology to Functional Genomics 245 207-222, 2002
View abstract

Barratt EA, Thompson GS, Harrison MA, Homans SW, Findlay JBC Structural analysis of Vma7p, subunit F of the vacuolar H+ATPase Biophysical Journal 82 2218-2218, 2002

Jones RPO, Hunt I, Jaeger J, Ward A, O'Reilly J, Barratt EA, Findlay JBC, Harrison MA Expression, purification and secondary structure analysis of Saccharomyces cerevisiae vacuolar membrane H + -ATPase subunit F (Vma7p) Molecular Membrane Biology 18 283-290, 2001
DOI:10.1080/09687680110099538

Harrison MA, Powell B, Finbow ME, Findlay JBC Identification of lipid-accessible sites on the Nephrops 16-kDa proteolipid incorporated into a hybrid H+-ATPase: Site directed labeling with N-(10pyrenyl)cyclohexylcarbodiimide and fluorescence quenching analysis Biochemistry 39 7531-7537, 2000

Harrison MA, Murray JM, Powell B, Kim YI, Finbow ME, Findlay JBC Helical interactions and membrane disposition of the 16-kDa proteolipid subunit of the vacuolar H+ ATPase analyzed by cysteine replacement mutagenesis Journal of Biological Chemistry 274 25461-25470, 1999

Murray JG, Powell B, Harrison MA, Findlay JBC Structure and function of the 16-kDa proteolipid of the yeast vacuolar H+ ATPase Acta Physiologica Scandinavica 163 311-311, 1998

Harrison MA, Finbow ME, Findlay JBC Postulate for the molecular mechanism of the vacuolar H+-ATPase (hypothesis) Molecular Membrane Biology 14 1-3, 1997

Findlay JBC, Finbow ME, Jones PC, Kim YI, Harrison MA, Hughes G A structure-based model for the 16 kDa membrane sector of the vacuolar H+-ATPase Biochemical Society Transactions 25 1107-1113, 1997

Finbow ME, Harrison MA The vacuolar H+-ATPase: A universal proton pump of eukaryotes Biochemical Journal 324 697-712, 1997

Powell B, Harrison MA, Findlay JBC Mutagenesis and chemical probing of the 16kDa proteolipid Faseb Journal 11 565-565, 1997

Hughes G, Harrison MA, Kim YI, Griffiths DE, Finbow ME, Findlay JBC Interaction of dibutyltin-3-hydroxyflavone bromide with the 16 kDa proteolipid indicates the disposition of proton translocation sites of the vacuolar H+-ATPase Biochemical Journal 317 425-431, 1996

Jones PC, Harrison MA, Kim YI, Finbow ME, Findlay JBC The first putative transmembrane helix of the 16 kDa proteolipid lines a pore in the V-0 sector of the vacuolar H+-ATPase Biochemical Journal 312 739-747, 1995
View abstract

Finbow ME, Harrison M, Jones P Ductin– a proton pump component, a gap junction channel and a neurotransmitter release channel BioEssays 17 247-255, 1995
DOI:10.1002/bies.950170311
View abstract

Nore BF, Harrison MA, Keen JN, Allen JF Partial-Purification of A Cyanobacterial Membrane-Protein with Amino-Terminal Sequence Similarity to the N-Methylphenylalanine Pilins Acta Chemica Scandinavica 48 578-581, 1994
View abstract

Harrison MA, Jones PC, Kim YI, Holzenburg A, Finbow ME, Findlay JBC Structure and Function of Related Proton Channel-Forming Proteins Pure and Applied Chemistry 66 35-41, 1994
View abstract

Harrison MA, Jones PC, Kim YI, Finbow ME, Findlay JBC Functional-Properties of A Hybrid Vacuolar H+-Atpase in Saccharomyces Cells Expressing the Nephrops 16-Kda Proteolipid European Journal of Biochemistry 221 111-120, 1994
View abstract

Jones PC, Harrison MA, Kim YI, Finbow ME, Findlay JBC Structure and Function of the Proton-Conducting Sector of the Vacuolar H+-Atpase Biochemical Society Transactions 22 805-809, 1994

Harrison MA, Allen JF Differential Phosphorylation of Individual Lhc-Ii Polypeptides During Short-Term and Long-Term Acclimation to Light Regime in the Green-Alga Dunaliella-Salina Biochimica et Biophysica Acta 1141 37-44, 1993
View abstract

Harrison MA, Nemson JA, Melis A Assembly and Composition of the Chlorophyll A-B Light-Harvesting Complex of Barley (Hordeum-Vulgare L) - Immunochemical Analysis of Chlorophyll B-Less and Chlorophyll B-Deficient Mutants Photosynthesis Research 38 141-151, 1993
View abstract

Melis A, Nemson JA, Harrison MA Damage to Functional Components and Partial Degradation of Photosystem-Ii Reaction Center Proteins Upon Chloroplast Exposure to Ultraviolet-B Radiation Biochimica et Biophysica Acta 1100 312-320, 1992
View abstract

Harrison MA, Melis A, Allen JF Restoration of Irradiance-Stressed Dunaliella-Salina (Green-Alga) to Physiological Growth-Conditions - Changes in Antenna Size and Composition of Photosystem-Ii Biochimica et Biophysica Acta 1100 83-91, 1992
View abstract

Harrison MA, Melis A Organization and Stability of Polypeptides Associated with the Chlorophyll A-B Light-Harvesting Complex of Photosystem-Ii Plant and Cell Physiology 33 627-637, 1992
View abstract

Nore BF, Harrison MA, Andersson BA, Allen JF Partial-Purification of the 15 Kda Phosphoprotein of Synechococcus 6301 Photosynthesis Research 34 223-223, 1992

Harrison MA, Allen JF Protein-Phosphorylation and Mg-2+ Influence Light Harvesting and Electron-Transport in Chloroplast Thylakoid Membrane Material Containing Only the Chlorophyll-A/B-Binding Light-Harvesting Complex of Photosystem-Ii and Photosystem-I European Journal of Biochemistry 204 1107-1114, 1992
View abstract

Harrison MA, Allen JF Light-Dependent Phosphorylation of Photosystem-Ii Polypeptides Maintains Electron-Transport at High Light-Intensity - Separation from Effects of Phosphorylation of Lhc-Ii Biochimica et Biophysica Acta 1058 289-296, 1991
View abstract

Tsinoremas NF, Castets AM, Harrison MA, Allen JF, Demarsac NT Photosynthetic Electron-Transport Controls Nitrogen Assimilation in Cyanobacteria by Means of Posttranslational Modification of the Glnb Gene-Product Proceedings Of The National Academy Of Sciences Of The United States Of America 88 4565-4569, 1991
View abstract

Harrison MA, Tsinoremas NF, Allen JF Cyanobacterial Thylakoid Membrane-Proteins Are Reversibly Phosphorylated Under Plastoquinone-Reducing Conditions Invitro Febs Letters 282 295-299, 1991
View abstract

Smith BM, Morrissey PJ, Guenther JE, Nemson JA, Harrison MA, Allen JF, Melis A Response of the Photosynthetic Apparatus in Dunaliella-Salina (Green-Algae) to Irradiance Stress Plant Physiology 93 1433-1440, 1990

Harrison MA, Keen JN, Findlay JBC, Allen JF Modification of A Glnb-Like Gene-Product by Photosynthetic Electron-Transport in the Cyanobacterium-Synechococcus 6301 Febs Letters 264 25-28, 1990

Allen JF, Harrison MA, Holmes NG Protein-Phosphorylation and Control of Excitation-Energy Transfer in Photosynthetic Purple Bacteria and Cyanobacteria Biochimie 71 1021-1028, 1989