TY - JOUR

T1 - Homology modelling of the GABA transporter and analysis of tiagabine binding

AU - Skovstrup, Søren

AU - Taboureau, Olivier

AU - Bräuner-Osborne, Hans

AU - Jørgensen, Flemming Steen

PY - 2010

Y1 - 2010

N2 - A homology model of the human GABA transporter (GAT-1) based on the recently reported crystal structures of the bacterial leucine transporter from Aquifex aeolicus (LeuT) was developed. The stability of the resulting model embedded in a membrane environment was analyzed by extensive molecular dynamics (MD) simulations. Based on docking studies and subsequent MD simulations of three compounds, the endogenous ligand GABA and two potent inhibitors, (R)-nipecotic acid and the anti-epilepsy drug tiagabine, various binding modes were identified and are discussed. Whereas GABA and (R)-nipecotic acid, which are both substrates, are stabilised with residues located deep inside the occluded state binding pocket (including residues Tyr 60 and Ser 396), tiagabine, which contains a large aliphatic side chain, is stabilised in a binding mode that extends from the substrate binding pocket (i.e., stabilised by Phe 294) to the extracellular vestibule, where the side chain is stabilised by aliphatic residues. The tiagabine binding mode, reaching from the substrate binding site to the extracellular vestibule, forces the side chain of Phe 294 to adopt a distinct conformation from that found in the occluded conformation of the transporter. Hence, in presence of tiagabine, GAT-1 is constrained in an open-to-out conformation. Our results may be of particular interest for the design of new GAT-1 inhibitors.

AB - A homology model of the human GABA transporter (GAT-1) based on the recently reported crystal structures of the bacterial leucine transporter from Aquifex aeolicus (LeuT) was developed. The stability of the resulting model embedded in a membrane environment was analyzed by extensive molecular dynamics (MD) simulations. Based on docking studies and subsequent MD simulations of three compounds, the endogenous ligand GABA and two potent inhibitors, (R)-nipecotic acid and the anti-epilepsy drug tiagabine, various binding modes were identified and are discussed. Whereas GABA and (R)-nipecotic acid, which are both substrates, are stabilised with residues located deep inside the occluded state binding pocket (including residues Tyr 60 and Ser 396), tiagabine, which contains a large aliphatic side chain, is stabilised in a binding mode that extends from the substrate binding pocket (i.e., stabilised by Phe 294) to the extracellular vestibule, where the side chain is stabilised by aliphatic residues. The tiagabine binding mode, reaching from the substrate binding site to the extracellular vestibule, forces the side chain of Phe 294 to adopt a distinct conformation from that found in the occluded conformation of the transporter. Hence, in presence of tiagabine, GAT-1 is constrained in an open-to-out conformation. Our results may be of particular interest for the design of new GAT-1 inhibitors.

KW - Former Faculty of Pharmaceutical Sciences

U2 - 10.1002/cmdc.201000100

DO - 10.1002/cmdc.201000100

M3 - Journal article

C2 - 20491137

VL - 5

SP - 986

EP - 1000

JO - Farmaco

JF - Farmaco

SN - 1860-7179

IS - 7

ER -