A single nucleotide polymorphism in the human serotonin transporter introduces a new site for N-linked glycosylation

Rasmussen, Trine N.; Plenge, Per; Bay, Tina; Egebjerg, Jan; Gether, Ulrik

doi:10.1016/j.neuropharm.2009.05.009

Cited by 23 publications

(22 citation statements)

References 33 publications

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“…CAD cells were transiently transfected with SERT tagged at the N terminus with a short c-Myc epitope, which does not affect expression and function of SERT (38). Cells were biotinylated with the reducible biotinylation reagent, sulfo-NHS-S-S-biotin, containing a cleavable disulfide bond, and subsequently internalization was allowed by incubation of the cells at 37°C for 2 h. Surface biotin was removed by treating cells with a reducing agent (2-mercaptoethanesulfonic acid (MesNa)), and intracellular biotinylated protein was isolated from the lysed cells with avidin beads, followed by SDS-PAGE and immunoblotting.…”

Section: Resultsmentioning

confidence: 99%

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The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation

Rahbek-Clemmensen

Bay

Eriksen

et al. 2014

Journal of Biological Chemistry

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Background: SERT is a target for antidepressants, but little is known about its constitutive cellular trafficking properties. Results: SERT undergoes marked constitutive internalization, and internalized SERT co-localizes primarily with markers of the late endosomal/lysosomal pathway. Conclusion: SERT is primarily sorted to degradation rather than to recycling. Significance: The findings are important for our general understanding of how SERT regulates serotonin signaling.

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Section: Resultsmentioning

confidence: 99%

“…Possibly, this reduction is the result of a structural perturbation caused by inserting the HA sequence between the two glycosylation sites in the EL2. Indeed, we know from previous studies that mutations interfering with glycosylation in EL2 of SERT alter transporter protein expression (38).…”

Section: Discussionmentioning

confidence: 99%

The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation

Rahbek-Clemmensen

Bay

Eriksen

et al. 2014

Journal of Biological Chemistry

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“…Human embryonic kidney 293 (HEK293) cells and COS-7 cells were maintained as previously described (70,71). Cath.-a-differentiated (CAD) cells from a catecholaminergic cell line were grown in media composed of 50% DMEM and 50% Ham's F-12 nutrient mixture (all from Invitrogen) at 37°C in a humidified incubator with 5% CO2.…”

Section: Methodsmentioning

confidence: 99%

Missense dopamine transporter mutations associate with adult parkinsonism and ADHD

Herborg

Skjørringe²,

Yasmeen³

et al. 2014

J. Clin. Invest.

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171

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Parkinsonism and attention deficit hyperactivity disorder (ADHD) are widespread brain disorders that involve disturbances of dopaminergic signaling. The sodium-coupled dopamine transporter (DAT) controls dopamine homeostasis, but its contribution to disease remains poorly understood. Here, we analyzed a cohort of patients with atypical movement disorder and identified 2 DAT coding variants, DAT-Ile312Phe and a presumed de novo mutant DAT-Asp421Asn, in an adult male with early-onset parkinsonism and ADHD. According to DAT single-photon emission computed tomography (DAT-SPECT) scans and a fluoro-deoxy-glucose-PET/MRI (FDG-PET/MRI) scan, the patient suffered from progressive dopaminergic neurodegeneration. In heterologous cells, both DAT variants exhibited markedly reduced dopamine uptake capacity but preserved membrane targeting, consistent with impaired catalytic activity. Computational simulations and uptake experiments suggested that the disrupted function of the DAT-Asp421Asn mutant is the result of compromised sodium binding, in agreement with Asp421 coordinating sodium at the second sodium site. For DAT-Asp421Asn, substrate efflux experiments revealed a constitutive, anomalous efflux of dopamine, and electrophysiological analyses identified a large cation leak that might further perturb dopaminergic neurotransmission. Our results link specific DAT missense mutations to neurodegenerative early-onset parkinsonism. Moreover, the neuropsychiatric comorbidity provides additional support for the idea that DAT missense mutations are an ADHD risk factor and suggests that complex DAT genotype and phenotype correlations contribute to different dopaminergic pathologies.

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“…Glycosylation. The SLC6 NTTs each possess two to four consensus sites for N-linked glycosylation (N-X-S/T) within EL2 that are subject to N-linked glycosylation (Table 6) (Tate and Blakely, 1994;Olivares et al, 1995;Bennett and Kanner, 1997;Torres et al, 2003a;Li et al, 2004;Rasmussen et al, 2009). Differences in the degree of glycosylation are observed among different tissue, brain regions, and cell types and during development (Lew et al, 1991;Patel et al, 1994;Qian et al, 1995;Nguyen and Amara, 1996).…”

Section: A Post-translational Modificationsmentioning

confidence: 99%

SLC6 Neurotransmitter Transporters: Structure, Function, and Regulation

et al. 2011

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A single nucleotide polymorphism in the human serotonin transporter introduces a new site for N-linked glycosylation

Cited by 23 publications

References 33 publications

The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation

The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation

Missense dopamine transporter mutations associate with adult parkinsonism and ADHD

SLC6 Neurotransmitter Transporters: Structure, Function, and Regulation

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