Effect of palmitoylation on the dimer formation of the human dopamine transporter

Zeppelin, Talia; Pedersen, Kasper B.; Berglund, Nils; Periole, Xavier; Schiøtt, Birgit

doi:10.1038/s41598-021-83374-y

Cited by 9 publications

(8 citation statements)

References 53 publications

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“…Among the five possible conserved Cys residues in rodents (r) and human (h), each (rDAT: 6,135,341,522,580;hDAT: 6,135,342,523 and 581) are exposed for palmitoylation, and mutational studies indicate 60% reduction in incorporation at the Cys580 (rDAT) site (Rastedt et al, 2017). Furthermore, Cys581 palmitoylation of hDAT leads to formation of stable and energetically favorable hDAT dimers with functional effects on DA uptake (Zeppelin et al, 2021). Various studies found that DAT palmitoylation increases the DA uptake Vmax and reduces the likelihood of degradation whereas depalmitoylation enhances DA efflux, decreases DA uptake Vmax, and increases the occurrence of lysosomal-associated degradation similar to the effects of phosphorylation (Khoshbouei et al, 2004;Moritz et al, 2015;Wang C. et al, 2016).…”

Section: Palmitoylationmentioning

confidence: 99%

“…Palmitoylation on TM12 has a favorable impact on oligomerization but the functional consequences on DAT are unclear (Zeppelin et al, 2021). Many attempts to understand the conformational dynamics of DAT oligomers and to predict distinct oligomeric interfaces have been made using MD simulations (Gur et al, 2017;Jayaraman et al, 2018;Zeppelin et al, 2021). Coarsegrained MD simulations of hDAT show that TM9, TM11, and/or TM12 provide the dimeric interface for oligomerization (Zeppelin et al, 2021).…”

Section: Dat Oligomerizationmentioning

confidence: 99%

“…Many attempts to understand the conformational dynamics of DAT oligomers and to predict distinct oligomeric interfaces have been made using MD simulations (Gur et al, 2017;Jayaraman et al, 2018;Zeppelin et al, 2021). Coarsegrained MD simulations of hDAT show that TM9, TM11, and/or TM12 provide the dimeric interface for oligomerization (Zeppelin et al, 2021). Unrestrained and unbiased coarse-grained MD simulation show that hDAT can form stable dimers through 6 different interfaces that include both symmetric and asymmetric dimers (Jayaraman et al, 2018).…”

Section: Dat Oligomerizationmentioning

confidence: 99%

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Overview of the structure and function of the dopamine transporter and its protein interactions

Nepal

Das²,

Reith³

et al. 2023

Front. Physiol.

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The dopamine transporter (DAT) plays an integral role in dopamine neurotransmission through the clearance of dopamine from the extracellular space. Dysregulation of DAT is central to the pathophysiology of numerous neuropsychiatric disorders and as such is an attractive therapeutic target. DAT belongs to the solute carrier family 6 (SLC6) class of Na+/Cl− dependent transporters that move various cargo into neurons against their concentration gradient. This review focuses on DAT (SCL6A3 protein) while extending the narrative to the closely related transporters for serotonin and norepinephrine where needed for comparison or functional relevance. Cloning and site-directed mutagenesis experiments provided early structural knowledge of DAT but our contemporary understanding was achieved through a combination of crystallization of the related bacterial transporter LeuT, homology modeling, and subsequently the crystallization of drosophila DAT. These seminal findings enabled a better understanding of the conformational states involved in the transport of substrate, subsequently aiding state-specific drug design. Post-translational modifications to DAT such as phosphorylation, palmitoylation, ubiquitination also influence the plasma membrane localization and kinetics. Substrates and drugs can interact with multiple sites within DAT including the primary S1 and S2 sites involved in dopamine binding and novel allosteric sites. Major research has centered around the question what determines the substrate and inhibitor selectivity of DAT in comparison to serotonin and norepinephrine transporters. DAT has been implicated in many neurological disorders and may play a role in the pathology of HIV and Parkinson’s disease via direct physical interaction with HIV-1 Tat and α-synuclein proteins respectively.

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Section: Palmitoylationmentioning

confidence: 99%

Section: Dat Oligomerizationmentioning

confidence: 99%

Section: Dat Oligomerizationmentioning

confidence: 99%

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Overview of the structure and function of the dopamine transporter and its protein interactions

Nepal

Das²,

Reith³

et al. 2023

Front. Physiol.

View full text Add to dashboard Cite

show abstract

“…1,12 As such, recent work has focused on elucidating the molecular mechanisms of post-translation modifications and its effect on human SLC6 transporters. 1,2 These studies include phosphorylation, 13−17 palmitoylation, 18,19 glycosylation, 20−24 and ubiquitination 25 and its implications on transporter dynamics, stability, oligomerization, trafficking, and uptake activity. The glycosylation of SLC6 transporters has been widely documented to affect transporter activity; 20−24 however, various mechanisms of how glycosylation mediates transporter function have been proposed for different SLC6 members.…”

Section: ■ Introductionmentioning

confidence: 99%

The Effects of N-Linked Glycosylation on SLC6 Transporters

Chan

Shukla

2023

J. Chem. Inf. Model.

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Membrane transporters of the solute carrier 6 (SLC6) family mediate various physiological processes by facilitating the translocation of amino acids, neurotransmitters, and other metabolites. In the body, the activity of these transporters is tightly controlled through various post-translational modifications with implications on protein expression, stability, membrane trafficking, and dynamics. While N-linked glycosylation is a universal regulatory mechanism among eukaryotes, a consistent mechanism of how glycosylation affects the SLC6 transporter family remains elusive. It is generally believed that glycans influence transporter stability and membrane trafficking; however, the role of glycosylation on transporter dynamics remains disputable, with differing conclusions among individual transporters across the SLC6 family. In this study, we collected over 1 ms of aggregated all-atom molecular dynamics (MD) simulation data to systematically identify the impact of N-glycans on SLC6 transporter dynamics. We modeled four human SLC6 transporters, the serotonin, dopamine, glycine, and B 0 AT1 transporters, by first simulating all possible combinations of a glycan attached to each glycosylation site followed by investigating the effect of larger, oligo-N-linked glycans to each transporter. The simulations reveal that glycosylation does not significantly affect the transporter structure but alters the dynamics of the glycosylated extracellular loop and surrounding regions. The structural consequences of glycosylation on the loop dynamics are further emphasized with larger glycan molecules attached. However, no apparent differences in ligand stability or movement of the gating helices were observed, and as such, the simulations suggest that glycosylation does not have a profound effect on conformational dynamics associated with substrate transport.

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Section: Introductionmentioning

confidence: 99%

The Effects of N-linked Glycosylation on SLC6 Transporters

Chan

Shukla

2022

Preprint

View full text Add to dashboard Cite

Membrane transporters of the solute carrier 6 (SLC6) family mediate various physiological processes by facilitating the translocation of amino acids, neurotransmitters, and other metabolites. In the human body, these transporters are tightly controlled through various post-translational modifications with implications on protein expression, stability, membrane trafficking, and dynamics. While N-linked glycosylation is a universal regulatory mechanism among eukaryotes, the exact molecular mechanism of how glycosylation affects the SLC6 transporter family. It is generally believed that glycans influence transporter stability and membrane trafficking, however, the role of glycosylation on transporter dynamics remains inconsistent, with differing conclusions among individual transporters across the SLC6 family. In this study, we collected over 1 millisecond of aggregated all-atom molecular dynamics (MD) simulation data to identify the impact of N-glycans of four human SLC6 transporters: the serotonin transporter, dopamine transporter, glycine transporter, and neutral amino acid transporter B0AT1. We designed our computational study by first simulating all possible combination of a glycan attached to each glycosylation sites followed by investigating the effect of larger, oligo-N-linked glycans to each transporter. Our simulations reveal that glycosylation does not significantly affect transporter structure, but alters the dynamics of the glycosylated extracellular loop. The structural consequences of glycosylation on the loop dynamics are further emphasized in the presence of larger glycan molecules. However, no apparent trend in ligand stability or movement of gating helices was observed. In all, the simulations suggest that glycosylation does not consistently affect transporter structure and dynamics among the collective SLC6 family and should be characterized at a per-transporter level to further elucidate the underlining mechanisms of in vivo regulation.

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Effect of palmitoylation on the dimer formation of the human dopamine transporter

Cited by 9 publications

References 53 publications

Overview of the structure and function of the dopamine transporter and its protein interactions

Overview of the structure and function of the dopamine transporter and its protein interactions

The Effects of N-Linked Glycosylation on SLC6 Transporters

The Effects of N-linked Glycosylation on SLC6 Transporters

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