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Analysis of Neurotransmitter Transporter Posttranslational Modifications: Phosphorylation and Palmitoylation

  • James D. Foster
  • Danielle E. Rastedt
  • Sathyavathi ChallaSivaKanaka
  • Roxanne A. VaughanEmail author
Protocol
Part of the Neuromethods book series (NM, volume 118)

Abstract

Posttranslational modification is a key mechanism by which proteins are regulated. Phosphorylation is one of the most prevalent mechanisms of regulation and fatty acid modification of proteins is becoming more widely appreciated as a regulatory mechanism for many proteins. The dopamine transporter (DAT) is an integral plasma membrane protein that mediates reuptake of dopamine from the extraneuronal space and is subject to several posttranslational modifications including phosphorylation and palmitoylation. DAT belongs to a family of closely related neurotransmitter transporters that includes carriers for serotonin and norepinephrine, and dysregulation of these transporters is hypothesized to contribute to many neuropsychiatric disorders such as depression, schizophrenia, attention-deficit hyperactivity disorder, and Parkinson disease. Neurotransmitter transporter phosphorylation and palmitoylation can be studied using various methods including metabolic labeling, phosphoamino acid analysis, peptide mapping, and acyl-biotinyl exchange. In this chapter, we discuss these methods that are employed in our laboratory for studying these modifications on DAT.

Key words

Metabolic labeling Phosphoamino acid analysis phosphopeptide mapping Phosphospecific antibodies 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • James D. Foster
    • 1
  • Danielle E. Rastedt
    • 1
  • Sathyavathi ChallaSivaKanaka
    • 1
  • Roxanne A. Vaughan
    • 1
    Email author
  1. 1.Department of Biochemistry and Molecular BiologyUniversity of North Dakota School of Medicine and Health SciencesGrand ForksUSA

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