title: The functional domains of dopamine transporter for cocaine analog, CFT binding.
abstract: Cocaine analogue, CFT (2beta-carbomethoxy-3beta-(4-fluorophenyl) tropane) binding to dopamine transporter (DAT) in different species is quite heterogeneous. CFT is scarcely detected in bovine DAT whereas it is conspicuous in humans. To examine the structural basis for this functional discrepancy, we analyzed transporter chimeras of these two DATs. The CFT binding activities are avid in all of the chimeric DATs of which both of the 3rd and the 6-8th transmembrane domain (TM) are composed of human DAT sequences. On the contrary, CFT binding activities were scarcely detected if either or both of two regions are replaced with bovine sequences. These findings indicate that the CFT binding absolutely requires human DAT sequences, at least, in the regions encompassing the 3rd and 6-8th transmembrane domain (TM), and that these regions might contribute to form the 3-dimensional pocket for CFT binding.
authors: Department of Biochemistry, College of Medicine, Hanyang University.
authors: Sang-Hun
authors: SH
authors: Lee
authors: Mi-Yoon
authors: MY
authors: Chang
authors: Dae-Joon
authors: DJ
authors: Jeon
authors: Dong-Yul
authors: DY
authors: Oh
authors: Hyeon
authors: H
authors: Son
authors: Chang-Ho
authors: CH
authors: Lee
authors: Young-Seek
authors: YS
authors: Lee
authors: Yong-Sung
authors: YS
authors: Lee
mesh_terms: D000818:Animals; D002417:Cattle; D003042:Cocaine / Q000031:analogs & derivatives* / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D006801:Humans; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D009419:Nerve Tissue Proteins; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D050483:Dopamine Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; C493214:SLC6A3 protein, human; C021279:(1R-(exo,exo))-3-(4-fluorophenyl)-8-methyl-8- azabicyclo(3.2.1)octane-2-carboxylic acid, methyl ester; D003042:Cocaine