Crystal structures of MdfA complexed with acetylcholine and inhibitor reserpine

Biophysics Reports - Tập 2 - Trang 78-85 - 2016
Ming Liu1, Jie Heng2, Yuan Gao2, Xianping Wang2
1College of Biotechnology, Tianjin University of Science and Technology, Tianjin, China
2National Laboratory of Macromolecules, National Center of Protein Science - Beijing, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China

Tóm tắt

The DHA12 family of transporters contains a number of prokaryotic and eukaryote membrane proteins. Some of these proteins share conserved sites intrinsic to substrate recognition, structural stabilization and conformational changes. For this study, we chose the MdfA transporter as a model DHA12 protein to study some general characteristics of the vesicular neurotransmitter transporters (VNTs), which all belong to the DHA12 family. Two crystal structures were produced for E. coli MdfA, one in complex with acetylcholine and the other with potential reserpine, which are substrate and inhibitor of VNTs, respectively. These structures show that the binding sites of these two molecules are different. The Ach-binding MfdA is mainly dependent on D34, while reserpine-binding site is more hydrophobic. Based on sequence alignment and homology modelling, we were able to provide mechanistic insights into the association between the inhibition and the conformational changes of these transporters.

Tài liệu tham khảo

Adler J, Bibi E (2004) Determinants of substrate recognition by the Escherichia coli multidrug transporter MdfA identified on both sides of the membrane. J Biol Chem 279:8957–8965 Bravo DT, Kolmakova NG, Parsons SM (2005) New transport assay demonstrates vesicular acetylcholine transporter has many alternative substrates. Neurochem Int 47:243–247 Chandrasekaran A, Ojeda AM, Kolmakova NG, Parsons SM (2006) Mutational and bioinformatics analysis of proline- and glycine-rich motifs in vesicular acetylcholine transporter. J Neurochem 98:1551–1559 Darchen F, Scherman D, Henry JP (1989) Reserpine binding to chromaffin granules suggests the existence of two conformations of the monoamine transporter. Biochemistry 28:1692–1697 Davis IW, Murray LW, Richardson JS, Richardson DC (2004) MOLPROBITY: structure validation and all-atom contact analysis for nucleic acids and their complexes. Nucleic Acids Res 32:W615–619 De Jesus M, Jin J, Guffanti AA, Krulwich TA (2005) Importance of the GP dipeptide of the antiporter motif and other membrane-embedded proline and glycine residues in tetracycline efflux protein Tet(L). Biochemistry 44:12896–12904 D’Rozario RS, Sansom MS (2008) Helix dynamics in a membrane transport protein: comparative simulations of the glycerol-3-phosphate transporter and its constituent helices. Mol Membr Biol 25:571–583 Edgar R, Bibi E (1997) MdfA, an Escherichia coli multidrug resistance protein with an extraordinarily broad spectrum of drug recognition. J Bacteriol 179:2274–2280 Edgar R, Bibi E (1999) A single membrane-embedded negative charge is critical for recognizing positively charged drugs by the Escherichia coli multidrug resistance protein MdfA. EMBO J 18:822–832 Emsley P, Cowtan K (2004) Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol Crystallogr 60:2126–2132 Erickson JD, Eiden LE, Hoffman BJ (1992) Expression cloning of a reserpine-sensitive vesicular monoamine transporter. Proc Natl Acad Sci USA 89:10993–10997 Erickson JD, Schafer MK, Bonner TI, Eiden LE, Weihe E (1996) Distinct pharmacological properties and distribution in neurons and endocrine cells of two isoforms of the human vesicular monoamine transporter. Proc Natl Acad Sci USA 93:5166–5171 Finn JP, Edwards RH (1997) Individual residues contribute to multiple differences in ligand recognition between vesicular monoamine transporters 1 and 2. J Biol Chem 272:16301–16307 Heng J, Zhao Y, Liu M, Liu Y, Fan J, Wang X, Zhao Y, Zhang XC (2015) Substrate-bound structure of the E. coli multidrug resistance transporter MdfA. Cell Res 25:1060–1073 Holler JG, Christensen SB, Slotved HC, Rasmussen HB, Guzman A, Olsen CE, Petersen B, Molgaard P (2012) Novel inhibitory activity of the Staphylococcus aureus NorA efflux pump by a kaempferol rhamnoside isolated from Persea lingue Nees. J Antimicrob Chemother 67:1138–1144 Huang Y, Lemieux MJ, Song J, Auer M, Wang DN (2003) Structure and mechanism of the glycerol-3-phosphate transporter from Escherichia coli. Science 301:616–620 Jiang D, Zhao Y, Wang X, Fan J, Heng J, Liu X, Feng W, Kang X, Huang B, Liu J, Zhang XC (2013) Structure of the YajR transporter suggests a transport mechanism based on the conserved motif A. Proc Natl Acad Sci USA 110:14664–14669 Khare P, Ojeda AM, Chandrasekaran A, Parsons SM (2010) Possible important pair of acidic residues in vesicular acetylcholine transporter. Biochemistry 49:3049–3059 Lawal HO, Krantz DE (2013) SLC18: vesicular neurotransmitter transporters for monoamines and acetylcholine. Mol Aspects Med 34:360–372 Luo J, Parsons SM (2010) Conformational propensities of peptides mimicking transmembrane Helix 5 and Motif C in wild-type and mutant vesicular acetylcholine transporters. ACS Chem Neurosci 1:381–390 Merickel A, Rosandich P, Peter D, Edwards RH (1995) Identification of residues involved in substrate recognition by a vesicular monoamine transporter. J Biol Chem 270:25798–25804 Otwinowski Z, Minor W (1997) Processing of X-ray diffraction data collected in oscillation mode. Method Enzymol 276:307–326 Parsons SM (2000) Transport mechanisms in acetylcholine and monoamine storage. FASEB J 14:2423–2434 Paul S, Alegre KO, Holdsworth SR, Rice M, Brown JA, McVeigh P, Kelly SM, Law CJ (2014) A single-component multidrug transporter of the major facilitator superfamily is part of a network that protects Escherichia coli from bile salt stress. Mol Microbiol 92:872–884 Paulsen IT, Brown MH, Skurray RA (1996) Proton-dependent multidrug efflux systems. Microbiol Rev 60:575–608 Preskorn SH (2007) The evolution of antipsychotic drug therapy: reserpine, chlorpromazine, and haloperidol. J Psychiatr Pract 13:253–257 Putman M, van Veen HW, Konings WN (2000) Molecular properties of bacterial multidrug transporters. Microbiol Mol Biol Rev 64:672–693 Ugolev Y, Segal T, Yaffe D, Gros Y, Schuldiner S (2013) Identification of conformationally sensitive residues essential for inhibition of vesicular monoamine transport by the noncompetitive inhibitor tetrabenazine. J Biol Chem 288:32160–32171 Vardy E, Arkin IT, Gottschalk KE, Kaback HR, Schuldiner S (2004) Structural conservation in the major facilitator superfamily as revealed by comparative modeling. Protein Sci 13:1832–1840 Yaffe D, Radestock S, Shuster Y, Forrest LR, Schuldiner S (2013) Identification of molecular hinge points mediating alternating access in the vesicular monoamine transporter VMAT2. Proc Natl Acad Sci USA 110:E1332–E1341 Yelin R, Schuldiner S (1995) The pharmacological profile of the vesicular monoamine transporter resembles that of multidrug transporters. FEBS Lett 377:201–207 Zhang XC, Zhao Y, Heng J, Jiang D (2015) Energy coupling mechanisms of MFS transporters. Protein Sci 24:1560–1579