Photoaffinity labeling of the mammalian dopamine transporter

Floyd R. Sallee, Evan L. Fogel, Eric Schwartz, Sung Mo Choi, Dennis P. Curran, Hyman B. Niznik

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

A high affinity (1-2 nM) radioiodinated, photoaffinity probe for the dopamine transporter, 1-(2-[bis-(4-fluorophenyl)-methoxylethyl)-4-(2-[4-azido-3-[125I]iodophenyl]ethyl)piperazine ([125I]FAPP) has been synthesized. Upon photolysis, [125I]FAPP incorporates into a striatal polypeptide of apparent M, 62000 as visualized by autoradiography following sodium dodecyl sulfate-PAGE. Photoincorporation of [125I]FAPP into the Mr 62000 polypeptide was stereoselectively inhibited by various dopamine uptake agents with a potency order typical of the dopamine transporter. The glycoprotein nature of the apparent M, 62000 polypeptide was assessed following specific exo- and endoglycosidase treatment. The dopamine transporter appears to be associated with complex-type oligosaccharides as indexed by its susceptibility to neuraminidase but not α-mannosidase digestion. Complete NLinked deglycosylation of the neuronal dopamine transporter with the endoglycosidase, glycopeptidase-F, increased the electrophoretic mobility of the 62 kDa polypeptide to apparent Mr 48000. [125I]FAPP should prove to be a useful probe for the molecular characterization of the dopamine uptake site in various tissues and under certain pathophysiological states. Dopamine uptake; Transporter; Photoaffinity labeling; Deglycosylation.

Original languageEnglish (US)
Pages (from-to)219-224
Number of pages6
JournalFEBS Letters
Volume256
Issue number1-2
DOIs
StatePublished - Oct 9 1989
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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    Sallee, F. R., Fogel, E. L., Schwartz, E., Choi, S. M., Curran, D. P., & Niznik, H. B. (1989). Photoaffinity labeling of the mammalian dopamine transporter. FEBS Letters, 256(1-2), 219-224. https://doi.org/10.1016/0014-5793(89)81752-1