Exploring the interaction of SV2A with racetams using homology modelling, molecular dynamics and site-directed mutagenesis. by Lee, J, Daniels, V, Sands, Z, Lebon, F, Shi, J, Biggin, P

Study of human Orexin-1 and -2 G-protein-coupled receptors with novel and published antagonists by modeling, molecular dynamics simulations, and site-directed mutagenesis. by Heifetz, A, Morris, G, Biggin, P, Barker, O, Fryatt, T, Bentley, J, Hallett, D, Manikowski, D, Pal, S, Reifegerste, R, Slack, M, Law, R

“…The class A G-protein-coupled receptors (GPCRs) Orexin-1 (OX1) and Orexin-2 (OX2) are located predominantly in the brain and are linked to a range of different physiological functions, including the control of feeding, energy metabolism, modulation of neuro-endocrine function, and regulation of the sleep-wake cycle. Site-directed mutagenesis (SDM) and domain exchange (chimera) studies have provided important insight into key features of the OX1 and OX2 binding sites. …”

Agonist and antagonist binding in human glycine receptors. by Yu, R, Hurdiss, E, Greiner, T, Lape, R, Sivilotti, L, Biggin, P

“…Thus, we constructed a model of the hGlyR and validated it against previously reported mutagenesis data. We used molecular dynamics to refine the model and to explore binding of both an agonist (glycine) and an antagonist (strychnine). …”

Homology modelling of human P-glycoprotein. by Domicevica, L, Biggin, P

“…Much of our knowledge to date has been derived from mutagenesis and assay data. However, in recent years, there has also been great progress in structural biology and although the structure of human P-gp has not yet been solved, there are now a handful of related structures on which homology models can be built to aid in the interpretation of the vast amount of experimental data that currently exists. …”

Chloride ions in the pore of glycine and GABA channels shape the time course and voltage dependence of agonist currents. by Moroni, M, Biro, I, Giugliano, M, Vijayan, R, Biggin, P, Beato, M, Sivilotti, L

“…This hypothesis is supported by the observation that the sensitivity of channel gating to intracellular chloride is abolished if the channel is engineered to become cation selective or if positive charges in the external pore vestibule are eliminated by mutagenesis. The appropriate interaction between permeating ions and channel pore is also necessary to maintain the channel voltage sensitivity of gating, which prolongs current decay at depolarized potentials. …”

Mutationmapper: a tool to aid the mapping of protein mutation data by Vohra, S, Biggin, P

“…There has been a rapid increase in the amount of mutational data due to, amongst other things, an increase in single nucleotide polymorphism (SNP) data and the use of site-directed mutagenesis as a tool to help dissect out functional properties of proteins. …”

The startle disease mutation E103K impairs activation of human homomeric α1 glycine receptors by disrupting an intersubunit salt bridge across the agonist binding site. by Safar, F, Hurdiss, E, Erotocritou, M, Greiner, T, Lape, R, Irvine, M, Fang, G, Jane, D, Yu, R, Dämgen, M, Biggin, P, Sivilotti, L

“…We investigated this hypothesis in recombinant human α1 GlyR by site-directed mutagenesis and functional measurements of agonist efficacy and potency by whole cell patch clamp and single channel recording. …”

Role of the Cys loop and transmembrane domain in the allosteric modulation of α4β2 nicotinic acetylcholine receptors by Alcaino, C, Musgaard, M, Minguez, T, Mazzaferro, S, Faundez, M, Iturriaga-Vasquez, P, Biggin, P, Bermudez, I

“…Here, using homology modelling in combination with mutagenesis and electrophysiology, we identified the binding site for potentiating dFBr on the top-half of a cavity between the third (M3) and fourth transmembrane (M4) α-helices of the α4 subunit. …”

Molecular determinants for competitive inhibition of alpha4beta2 nicotinic acetylcholine receptors. by Iturriaga-Vásquez, P, Carbone, A, García-Beltrán, O, Livingstone, P, Biggin, P, Cassels, B, Wonnacott, S, Zapata-Torres, G, Bermudez, I

“…We addressed this issue by examining the effects of DHbetaE and a range of aromatic Erythrina alkaloids on [(3)H]cytisine binding and receptor function in conjunction with homology models of the alpha4beta2 nAChR, mutagenesis, and functional assays. The lactone group of DHbetaE and a hydroxyl group at position C-16 in aromatic Erythrina alkaloids were identified as major determinants of potency, which was decreased when the conserved residue Tyr126 in loop A of the alpha4 subunit was substituted by alanine. …”

Additional acetylcholine (ACh) binding site at alpha4/alpha4 interface of (alpha4beta2)2alpha4 nicotinic receptor influences agonist sensitivity. by Mazzaferro, S, Benallegue, N, Carbone, A, Gasparri, F, Vijayan, R, Biggin, P, Moroni, M, Bermudez, I

“…Using fully concatenated (α4β2)(2)α4 nAChRs in conjunction with structural modeling, chimeric receptors, and functional mutagenesis, we have identified an additional site at the α4(+)/α4(-) interface that accounts for isoform-specific agonist sensitivity of the (α4β2)(2)α4 nAChR. …”

Additional Acetylcholine (ACh) Binding Site at alpha 4/alpha 4 Interface of (alpha 4 beta 2)(2)alpha 4 Nicotinic Receptor Influences Agonist Sensitivity by Mazzaferro, S, Benallegue, N, Carbone, A, Gasparri, F, Vijayan, R, Biggin, P, Moroni, M, Bermudez, I

“…Using fully concatenated (α4β2) 2α4 nAChRs in conjunction with structural modeling, chimeric receptors, and functional mutagenesis, we have identified an additional site at the α4(+)/α4(- ) interface that accounts for isoform-specific agonist sensitivity of the (α4β2) 2α4 nAChR. …”

Structural, functional and computational studies on the SLC7 family of amino acid transporters by Kolokouris, D

“…Structural analyses and mutagenesis suggested the root of the homologue discrepancy to be two bulky, hydrophobic residues absent from AtCAT4 and other CATs. …”

Allosteric contributions of muscle nicotinic acetylcholine receptor residues in small-molecule interactions, disease and subunit assembly by Epstein, M

“…In the first part of this project, we sought to rationalise the structure activity relationship data of a series of congeneric BPDs using molecular dynamics (MD) simulations, followed by mutagenesis and electrophysiology to elucidate molecular determinants for their interactions.…”