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Photoaffinity labelling of dopamine D2receptors by [3H]azidomethylspiperone

Authors
Journal
FEBS Letters
0014-5793
Publisher
Wiley Blackwell (John Wiley & Sons)
Publication Date
Volume
209
Issue
1
Identifiers
DOI: 10.1016/0014-5793(86)81086-9
Keywords
  • D 2 Dopamine Receptorphotoaffinity Labelinglectin Affinity Chromatographysds-Pagefluorography

Abstract

Abstract We have characterized the dopamine D 2 receptor photaffinity probe, [ 3H]azido- N-methylspiperone ([ 3H]AMS). In the absence of light, [ 3H]AMS bound reversibly and with high affinity ( K d 70 pM) to sites in canine striatal membranes and was competitively inhibited by dopaminergic agonists and antagonists with an appropriate D 2 receptor specificity. Upon photolysis, [ 3H]AMS covalently incorporated into a peptide of M r 92000 as assessed by fluorography following SDS-polyacrylamide gel electrophoresis. Labelling of this peptide was specifically and stereoselectively blocked by D 2 antagonists and agonists. Minor specifically labelled peptides of M r 70000-55000 were observed under some conditions and were the result of proteolytic degradation of the peptide at M r, 92000.

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