Research School of Biomedical Sciences
Integrative Membrane Biology Group
Integrative Membrane Biology (IMB) is a leading UK centre for the study of membranes, with strong international links and a diversity of expertise ranging from membrane protein structure, through cell biology & physiology, to therapeutics.
Its integrative approach is facilitated by the co-location of neuroscientists, physiologists and pharmacologists with researchers in membrane protein structure. In addition, very strong interdisciplinary links are maintained with researchers in Medicine, Chemistry and Physics .
Cutting-edge infrastructure developed by IMB includes a Bioimaging Facility, with TIRF/swept field and other confocal microscopes, and new laboratories for both robotic high-throughput membrane protein expression and functional analysis using fluorescence technology and planar patch-clamp.
Some Research Highlights:
Li, J., Cubbon, R.M., Wilson, L.A., Amer, M.S., McKeown, L., Hou, B., Majeed, Y., Tumova, S., Seymour, V.A.L., Taylor, H., Stacey, M., O’Regan, D., Foster, R., Porter, K.E., Kearney, M.T., Beech, D.J. (2011). Orai1 and CRAC channel dependence of VEGF-activated Ca2+-entry and endothelial tube formation. Circulation Research Published On-line.
Huysmans, GH; Baldwin, SA; Brockwell, DJ; Radford, SE The transition state for folding of an outer membrane protein. Proc Natl Acad Sci U S A 107 4099-4104, 2010. DOI:10.1073/pnas.0911904107
Newstead S, Drew D, Cameron AD, Postis VL, Xia X, Fowler PW, Ingram JC, Carpenter EP, Sansom MS, McPherson MJ, Baldwin SA, Iwata S. Crystal structure of a prokaryotic homologue of the mammalian oligopeptide-proton symporters, PepT1 and PepT2. EMBO J. 2010 Dec 3. [Epub ahead of print]
Cheong, A; Li, J; Sukumar, P; Kumar, B; Zeng, F; Riches, K; Munsch, C; Wood, IC; Porter, KE; Beech, DJ Potent suppression of vascular smooth muscle cell migration and human neointimal hyperplasia by KV1.3 channel blockers. Cardiovasc Res -, 2010. DOI:10.1093/cvr/cvq305
Naylor, J; Li, J; Milligan, CJ; Zeng, F; Sukumar, P; Hou, B; Sedo, A; Yuldasheva, N; Majeed, Y; Beri, D; Jiang, S; Seymour, VA; McKeown, L; Kumar, B; Harteneck, C; O'Regan, D; Wheatcroft, SB; Kearney, MT; Jones, C; Porter, KE; Beech, DJ Pregnenolone sulphate- and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction. Circ Res 106 1507-1515, 2010. DOI:10.1161/CIRCRESAHA.110.219329
Xu, SZ; Sukumar, P; Zeng, F; Li, J; Jairaman, A; English, A; Naylor, J; Ciurtin, C; Majeed, Y; Milligan, CJ; Bahnasi, YM; Al-Shawaf, E; Porter, KE; Jiang, LH; Emery, P; Sivaprasadarao, A; Beech, DJ TRPC channel activation by extracellular thioredoxin. Nature 451 69-72, 2008. DOI:10.1038/nature06414
Clapcote, SJ; Duffy, S; Xie, G; Kirshenbaum, G; Bechard, AR; Rodacker Schack, V; Petersen, J; Sinai, L; Saab, BJ; Lerch, JP; Minassian, BA; Ackerley, CA; Sled, JG; Cortez, MA; Henderson, JT; Vilsen, B; Roder, JC Mutation I810N in the alpha3 isoform of Na+,K+-ATPase causes impairments in the sodium pump and hyperexcitability in the CNS. Proc Natl Acad Sci U S A 106 14085-14090, 2009. DOI:10.1073/pnas.0904817106
This finding highlights the critical importance of the a3 isoform of the Na+,K+-ATPase in the control of epileptiform activity and seizure behaviour. The publication of these findings in PNAS attracted significant interest from mainstream news media and was reported in New Scientist magazine.
Mann, RJ; Nasr, NE; Sinfield, JK; Paci, E; Donnelly, D The major determinant of Exendin-4/GLP-1 differential affinity at the rat GLP-1 receptor N-terminal domain is a hydrogen bond from Ser-32 of exendin-4. Brit. J. Pharmacolol. 160 1973-1984, 2010
DOI:10.1111/j.1476-5381.2010.00834.x
The anti-diabetic drug exenatide binds to the isolated N-terminal domain of GLP-1R with much higher affinity than the natural hormone GLP-1. Using observed differences between the pharmacology of these peptides at human and rat receptors, this paper presents a model to explain peptide binding at GLP-1R.
Miller, PS; Barwell, J; Poyner, DR; Wigglesworth, MJ; Garland, SL; Donnelly, D Non-peptidic antagonists of the CGRP receptor, BIBN4096BS and MK-0974, interact with the calcitonin receptor-like receptor via methionine-42 and RAMP1 via tryptophan-74. Biochem Biophys Res Commun 391 437-442, 2010
DOI:10.1016/j.bbrc.2009.11.076
This work was a collaboration with GSK in which we used a molecular pharmacological approach to determine the first known contact residue on the GPCR component of the CGRP receptor with two non-peptidic antagonists which are in clinical trials as potential anti-migraine therapies. The subsequent crystal structure of these antagonists with the isolated N-terminal domains of the receptor confirmed the data.
Mann, RJ; Al-Sabah, S; de Maturana, RL; Sinfield, JK; Donnelly, D Functional coupling of Cys-226 and Cys-296 in the glucagon-like peptide-1 (GLP-1) receptor indicates a disulfide bond that is close to the activation pocket. Peptides 31 2289-2293, 2010
DOI:10.1016/j.peptides.2010.09.015
While there is no significant sequence conservation between Family A and Family B GPCRs, there are two conserved cysteine residues in Family B receptors which may reflect the presence of a disulphide bond analogous to that found in most Family A receptors. In this paper we demonstrate that these cysteines are functionally linked, suggesting that the disulphide exists. Furthermore, we demonstrated that the second extracellular loop of the receptor is important for receptor activity.
Mann, R; Wigglesworth, MJ; Donnelly, D Ligand-receptor interactions at the parathyroid hormone receptors: subtype binding selectivity is mediated via an interaction between residue 23 on the ligand and residue 41 on the receptor. Mol Pharmacol 74 605-613, 2008
DOI:10.1124/mol.108.048017
A collaboration with GSK in which a molecular pharmacological approach was used to determine a “rotamer toggle switch” mechanism by which parathyroid hormone receptors can select between different peptide ligands. The authors of a subsequent crystal paper which confirmed the model cited the work saying “This Leu-41 rotamer toggle switch mechanism was predicted in an elegant study by Donnelly and co-workers”.
Liu, B; Linley, JE; Du, X; Zhang, X; Ooi, L; Zhang, H; Gamper, N. The acute nociceptive signals induced by bradykinin in rat sensory neurons are mediated by inhibition of M-type K+ channels and activation of Ca2+-activated Cl- channels. J Clin Invest 120 1240-1252, 2010. DOI:10.1172/JCI41084
This study discovers novel signalling pathway underlying acute excitatory action of inflammatory mediators (such as bradykinin) on peripheral sensory neurons and, thus, provides a new mechanism for the acute inflammatory pain. The study demonstrated for the first time the role of Ca2+ activated Cl- channel Tmem16a (ANO1) in nociception and suggested two new targets for the inflammatory pain therapy.
Muench, SP; Huss, M; Song, CF; Phillips, C; Wieczorek, H; Trinick, J; Harrison, M Cryo-electron microscopy of the vacuolar ATPase motor reveals its mechanical and regulatory complexity Journal of Molecular Biology 386 989-999, 2009. DOI:10.1016/j.jmb.2009.01.014
Shimamura, T; Weyand, S; Beckstein, O; Rutherford, NG; Hadden, JM; Sharples, D; Sansom, MSP; Iwata, S; Henderson, PJF; Cameron, AD. Molecular Basis of Alternating Access Membrane Transport by the Sodium-Hydantoin Transporter Mhp1 SCIENCE 328 470-473, 2010. DOI:10.1126/science.1186303
This article reports the first example of an ion-coupled transporter with crystal structures of both outward- (periplasm) and inward- (cytoplasm) facing conformations.
Weyand, S; Shimamura, T; Yajima, S; Suzuki, S; Mirza, O; Krusong, K; Carpenter, EP; Rutherford, NG; Hadden, JM; O'Reilly, J; Ma, P; Saidijam, M; Patching, SG; Hope, RJ; Norbertczak, HT; Roach, PCJ; Iwata, S; Henderson, PJF; Cameron, AD. Structure and Molecular Mechanism of a Nucleobase-Cation-Symport-1 Family Transporter SCIENCE 322 709-713, 2008. DOI:10.1126/science.1164440
This study reports the structure of outward-facing Mhp1 conformations, offering fascinating insights into the transport mechanism.
Weiss, S. A., Bushby, R. J., Evans, S. D., Henderson, P. J. F., Jeuken, L. J. C. Characterisation of cytochrome bo3 activity in a native-like surface-tethered membrane,Biochem. J.,417, 555-560, 2009. DOI:10.1042/BJ20081345
This paper describes a new method to study redox-active membrane enzymes, especially quinone converting enzymes, in detergent free solutions.
Smith AJ & Lippiat JD. Voltage-dependent charge movement associated with activation of the CLC-5 2Cl-/1H+ exchanger. FASEB J 24, 3696-705, 2010.
First description of gating currents from the voltage-gated chloride channel family.
Smith, AJ; Lippiat, JD. Direct endosomal acidification by the outwardly rectifying CLC-5 Cl(-)/H(+) exchanger. J Physiol 588 2033-2045, 2010
DOI:10.1113/jphysiol.2010.188540.
This is the first demonstration that CLCs that act as chloride/proton exchange transporters can directly acidify intracellular vesicles by exchanging luminal chloride for protons. This function is impaired in Dent's disease in man.
Taneja, TK; Mankouri, J; Karnik, R; Kannan, S; Smith, AJ; Munsey, T; Christesen, HB; Beech, DJ; Sivaprasadarao, A Sar1-GTPase-dependent ER exit of KATP channels revealed by a mutation causing congenital hyperinsulinism. Hum Mol Genet 18 2400-2413, 2009. DOI:10.1093/hmg/ddp179
This study provides the cell biological basis of congenital hyperinsulinism and reports, for the first time, that a mutation in an ER exit motif can cause severe disease in humans.
Neale, EJ; Rong, H; Cockcroft, CJ; Sivaprasadarao, A. Mapping the Membrane-aqueous Border for the Voltage-sensing Domain of a Potassium Channel. Journal of Biological Chemistry 282 37597-37604, 2007
Using a protein chemistry approach, this study defines water crevices in the membrane embedded portion of the voltage sensor of a Kv channel, which play significant roles in for voltage sensing.