title: Structural determinants of species-selective substrate recognition in human and Drosophila serotonin transporters revealed through computational docking studies.
abstract: To identify potential determinants of substrate selectivity in serotonin (5-HT) transporters (SERT), models of human and Drosophila serotonin transporters (hSERT, dSERT) were built based on the leucine transporter (LeuT(Aa)) structure reported by Yamashita et al. (Nature 2005;437:215-223), PBDID 2A65. Although the overall amino acid identity between SERTs and the LeuT(Aa) is only 17%, it increases to above 50% in the first shell of the putative 5-HT binding site, allowing de novo computational docking of tryptamine derivatives in atomic detail. Comparison of hSERT and dSERT complexed with substrates pinpoints likely structural determinants for substrate binding. Forgoing the use of experimental transport and binding data of tryptamine derivatives for construction of these models enables us to critically assess and validate their predictive power: A single 5-HT binding mode was identified that retains the amine placement observed in the LeuT(Aa) structure, matches site-directed mutagenesis and substituted cysteine accessibility method (SCAM) data, complies with support vector machine derived relations activity relations, and predicts computational binding energies for 5-HT analogs with a significant correlation coefficient (R = 0.72). This binding mode places 5-HT deep in the binding pocket of the SERT with the 5-position near residue hSERT A169/dSERT D164 in transmembrane helix 3, the indole nitrogen next to residue Y176/Y171, and the ethylamine tail under residues F335/F327 and S336/S328 within 4 A of residue D98. Our studies identify a number of potential contacts whose contribution to substrate binding and transport was previously unsuspected.
authors.affiliation: Department of Chemistry, Vanderbilt University, Nashville, Tennessee 37235-1822, USA.
authors.forename: Kristian W
authors.initials: KW
authors.lastname: Kaufmann
authors.forename: Eric S
authors.initials: ES
authors.lastname: Dawson
authors.forename: L Keith
authors.initials: LK
authors.lastname: Henry
authors.forename: Julie R
authors.initials: JR
authors.lastname: Field
authors.forename: Randy D
authors.initials: RD
authors.lastname: Blakely
authors.forename: Jens
authors.initials: J
authors.lastname: Meiler
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003198:Computer Simulation; D004330:Drosophila / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006860:Hydrogen Bonding; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D021281:Quantitative Structure-Activity Relationship; D016415:Sequence Alignment; D012701:Serotonin / Q000031:analogs & derivatives / Q000737:chemistry; D050486:Serotonin Plasma Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity; D014363:Tryptamines / Q000737:chemistry
chemical_list: D029721:Drosophila Proteins; D008024:Ligands; D050486:Serotonin Plasma Membrane Transport Proteins; D014363:Tryptamines; D012701:Serotonin; C030820:tryptamine