The Importance of Company: Na+ and Cl− Influence Substrate Interaction with SLC6 Transporters and Other Proteins

Reith, Maarten E. A.; Zhen, Juan; Chen, N.

doi:10.1007/3-540-29784-7_4

Cited by 8 publications

(6 citation statements)

References 73 publications

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“…Reith et al (2006) suggest that the DAT in an outward facing conformational state is poised for binding Na ϩ , which in turn facilitates binding of cocaine analogs. Accordingly, the potential of JHW 007 to shift the DAT to an inward facing conformational equilibrium (Loland et al, 2008) Madras et al, 1989; but see Reith and Selmeci, 1992).…”

Section: Discussionmentioning

confidence: 99%

Dopamine Transporter-Dependent and -Independent Striatal Binding of the Benztropine Analog JHW 007, a Cocaine Antagonist with Low Abuse Liability

Kopajtic¹,

Liu²,

Surratt³

et al. 2010

J Pharmacol Exp Ther

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Section: Discussionmentioning

confidence: 99%

Dopamine Transporter-Dependent and -Independent Striatal Binding of the Benztropine Analog JHW 007, a Cocaine Antagonist with Low Abuse Liability

Kopajtic¹,

Liu²,

Surratt³

et al. 2010

J Pharmacol Exp Ther

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“…Follow-up validation studies included dose response experiments using the same assay, where 9 of these 15 compounds showed a dose-dependent effect for rescuing the lethality associated with the Fmr1(Ϫ/Ϫ) flies. It is noteworthy that three of these were related to the GABA signaling pathway, which is a major pathway underlying many of the symptoms of FXS (Krogsgaard-Larsen et al, 2000;Reith et al, 2006;Chang et al, 2008). These hits were GABA itself, nipecotic acid (a GABA reuptake inhibitor), and creatinine (a potential activator of the GABA A receptor).…”

Section: Successful Examples Of D Melanogaster In the Drug Discovmentioning

confidence: 99%

Human Disease Models inDrosophila melanogasterand the Role of the Fly in Therapeutic Drug Discovery

2011

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“…This family is characterized by an absolute requirement for Na + cotransport, which provides part of the driving force to accumulate neurotransmitter against an otherwise unfavorable concentration gradient. Transporters in this family also exhibit a variable and incompletely understood necessity for Cl − cotransport in the transport process ( Lester et al, 1996 ; Rudnick, 2002 ; Kanner, 2006 ; Reith et al, 2006 ).…”

Section: Introductionmentioning

confidence: 99%

GABA transporter function, oligomerization state, and anchoring: correlates with subcellularly resolved FRET

Moss

Imoukhuede

Scott

et al. 2009

Journal of General Physiology

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The mouse γ-aminobutyric acid (GABA) transporter mGAT1 was expressed in neuroblastoma 2a cells. 19 mGAT1 designs incorporating fluorescent proteins were functionally characterized by [3H]GABA uptake in assays that responded to several experimental variables, including the mutations and pharmacological manipulation of the cytoskeleton. Oligomerization and subsequent trafficking of mGAT1 were studied in several subcellular regions of live cells using localized fluorescence, acceptor photobleach Förster resonance energy transfer (FRET), and pixel-by-pixel analysis of normalized FRET (NFRET) images. Nine constructs were functionally indistinguishable from wild-type mGAT1 and provided information about normal mGAT1 assembly and trafficking. The remainder had compromised [3H]GABA uptake due to observable oligomerization and/or trafficking deficits; the data help to determine regions of mGAT1 sequence involved in these processes. Acceptor photobleach FRET detected mGAT1 oligomerization, but richer information was obtained from analyzing the distribution of all-pixel NFRET amplitudes. We also analyzed such distributions restricted to cellular subregions. Distributions were fit to either two or three Gaussian components. Two of the components, present for all mGAT1 constructs that oligomerized, may represent dimers and high-order oligomers (probably tetramers), respectively. Only wild-type functioning constructs displayed three components; the additional component apparently had the highest mean NFRET amplitude. Near the cell periphery, wild-type functioning constructs displayed the highest NFRET. In this subregion, the highest NFRET component represented ∼30% of all pixels, similar to the percentage of mGAT1 from the acutely recycling pool resident in the plasma membrane in the basal state. Blocking the mGAT1 C terminus postsynaptic density 95/discs large/zona occludens 1 (PDZ)-interacting domain abolished the highest amplitude component from the NFRET distributions. Disrupting the actin cytoskeleton in cells expressing wild-type functioning transporters moved the highest amplitude component from the cell periphery to perinuclear regions. Thus, pixel-by-pixel NFRET analysis resolved three distinct forms of GAT1: dimers, high-order oligomers, and transporters associated via PDZ-mediated interactions with the actin cytoskeleton and/or with the exocyst.

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The Importance of Company: Na+ and Cl− Influence Substrate Interaction with SLC6 Transporters and Other Proteins

Cited by 8 publications

References 73 publications

Dopamine Transporter-Dependent and -Independent Striatal Binding of the Benztropine Analog JHW 007, a Cocaine Antagonist with Low Abuse Liability

Dopamine Transporter-Dependent and -Independent Striatal Binding of the Benztropine Analog JHW 007, a Cocaine Antagonist with Low Abuse Liability

Human Disease Models inDrosophila melanogasterand the Role of the Fly in Therapeutic Drug Discovery

GABA transporter function, oligomerization state, and anchoring: correlates with subcellularly resolved FRET

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