Photoaffinity labeling of the mammalian dopamine transporter

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

Research output: Contribution to journalArticle

37 Citations (Scopus)

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

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Dopamine Plasma Membrane Transport Proteins
Labeling
Glycoside Hydrolases
Dopamine
Peptides
Mannosidases
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Corpus Striatum
Molecular Probes
Electrophoretic mobility
Dopamine Agents
Photolysis
Neuraminidase
Autoradiography
Oligosaccharides
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Digestion
Glycoproteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Sallee, F. R., Fogel, E., 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

Photoaffinity labeling of the mammalian dopamine transporter. / Sallee, Floyd R.; Fogel, Evan; Schwartz, Eric; Choi, Sung Mo; Curran, Dennis P.; Niznik, Hyman B.

In: FEBS Letters, Vol. 256, No. 1-2, 09.10.1989, p. 219-224.

Research output: Contribution to journalArticle

Sallee, FR, Fogel, E, Schwartz, E, Choi, SM, Curran, DP & Niznik, HB 1989, 'Photoaffinity labeling of the mammalian dopamine transporter', FEBS Letters, vol. 256, no. 1-2, pp. 219-224. https://doi.org/10.1016/0014-5793(89)81752-1
Sallee FR, Fogel E, Schwartz E, Choi SM, Curran DP, Niznik HB. Photoaffinity labeling of the mammalian dopamine transporter. FEBS Letters. 1989 Oct 9;256(1-2):219-224. https://doi.org/10.1016/0014-5793(89)81752-1
Sallee, Floyd R. ; Fogel, Evan ; Schwartz, Eric ; Choi, Sung Mo ; Curran, Dennis P. ; Niznik, Hyman B. / Photoaffinity labeling of the mammalian dopamine transporter. In: FEBS Letters. 1989 ; Vol. 256, No. 1-2. pp. 219-224.
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