N-Terminal Truncation of DAT Impairs AMPH-Induced DA Efflux

<p>Cells were preloaded with 15 μM DA and superfused with AMPH at concentrations ranging from 1 to 100 μM. AMPH-induced DA efflux was defined as the amount of DA released in response to the given concentration of AMPH minus the baseline value. Baseline DA release did not differ between FLAG-HA-DAT and FLAG-DAT (13.2 ± 2.9 and 10.2 ± 1.8, respectively; <i>n =</i> 18). The V_max of efflux was 31.1 ± 4.6 and 128.3 ± 12.0 pmol/mg protein/fraction (F(2,27) = 52.6, <i>p</i> < 0.0001) with a K_m for amphetamine of 7.8 ± 4.1 and 7.6 ± 2.2 μM, for FLAG-HA-DAT and FLAG-DAT, respectively (<i>n</i> = 4). For [³H]DA uptake, the V_max was 15.4 ± 2.5 and 18.3 ± 2.2 pmol/min/mg protein with a K_m of 1.2 ± 0.8 and 1.1 ± 0.4 μM for FLAG-HA-DAT and FLAG-DAT, respectively (F(2,49) = 1.78, <i>p</i> > 0.17).</p

N-Terminal Truncation of DAT Reduces AMPH-Induced Currents and DA Efflux

<div><p>Cells were voltage clamped with a whole-cell patch pipette while an amperometric electrode was placed onto the cell membrane. The internal solution of the whole-cell patch pipette contained 2 mM DA.</p> <p>(A) Representative trace of AMPH-induced whole-cell current obtained from FLAG-DAT cells upon AMPH (10 μM) bath application. The membrane potential of the cell was stepped to +100mV from a holding potential of –20 mV.</p> <p>(B) Oxidation current acquired concomitantly to the whole-cell current represented in panel A.</p> <p>(C and D) Representative current traces (whole-cell and amperometric, respectively) obtained from FLAG-del22-DAT cells using the same experimental protocol as in (A) and (B).</p></div

Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base-2

<p><b>Copyright information:</b></p><p>Taken from "Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base"</p><p>http://www.biomedcentral.com/1471-2105/8/177</p><p>BMC Bioinformatics 2007;8():177-177.</p><p>Published online 30 May 2007</p><p>PMCID:PMC1904246.</p><p></p> protomers oligomerize. There are three types of phenotypic change that are described by the ontology: changes in internalization, changes in signaling, and differences in the ligand binding of the oligomer as compared to any of the constituent protomers. The effect that ligand(s) binding to one or more of the protomers in an oligomer may have on binding of ligands to other protomers, or on the change in signaling, is described by the CrossTalk concept. The Internalization concept is used to describe changes that different ligands have on the trafficking of the Oligomer to the cell membrane. Any information that is available about the mechanism of activation of the Oligomer is stored in the MechanismOfActivation concept. The PhysiologicalRelevance concept stores information about the Oligomer

Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base-0

<p><b>Copyright information:</b></p><p>Taken from "Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base"</p><p>http://www.biomedcentral.com/1471-2105/8/177</p><p>BMC Bioinformatics 2007;8():177-177.</p><p>Published online 30 May 2007</p><p>PMCID:PMC1904246.</p><p></p> for each concept are listed in the middle section of the box. Arrows represent relationships between concepts, and open-ended arrows indicate "is a" relationships. The relationship of one concept to another is indicated by the text on each arrow. Arrows with [0..*] (zero or more) or [1..*] (one or more) indicate the number of instances of the concept at the end of the arrow that is associated with the concept at the beginning of the arrow. The Oligomer concept is central to the GPCR oligomerization ontology and all other concepts in the ontology relate to it either directly (e.g., Oligomer ''is described by'' IdentificationStudy) or indirectly (e.g., Oligomer ''is composed of'' one or more [1..*] Protein that ''belongs to'' Family)

Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base-1

<p><b>Copyright information:</b></p><p>Taken from "Requirements and ontology for a G protein-coupled receptor oligomerization knowledge base"</p><p>http://www.biomedcentral.com/1471-2105/8/177</p><p>BMC Bioinformatics 2007;8():177-177.</p><p>Published online 30 May 2007</p><p>PMCID:PMC1904246.</p><p></p>olecularStructure concept. Each MolecularStructure is created with an instance of MethodType, and may be analyzed by many computational methods (instances of Analysis). MethodType has two subclasses: IdentificationMethod, which is used to identify the oligomer, and CreationMethod, which is used to create the MolecularStructure. IdentificationMethod and CreationMethod can have many sub-concepts that describe the precise type of method. In this figure we show only a few examples of such concepts