Purification of the surface cAMP receptor in Dictyostelium

P. Klein, B. Knox, J. Borleis, P. Devreotes

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


We have previously identified and demonstrated reversible ligand-induced modification of the major cell surface cAMP receptor in Dictyostelium discoideum. The receptor, or a subunit of it, has been purified to homogeneity by hydroxylapatite chromatography followed by two-dimensional preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purification was monitored by following 32P(i) incorporated by photoaffinity labeling with 8-azido-[32P]cAMP or by in vivo labeling with 32P(i). Two interconvertible forms of the receptor, designated R (M(r) 40,000) and D (M(r) 43,000), co-purified. Two-dimensional peptide maps of independently purified and 125I-iodinated R and D forms of the receptor were nearly identical but did have several distinct peptides. The estimated 6000-fold purification required is consistent with the number of cell surface binding sites assuming there are not multiple binding sites/polypeptide. In the accompanying article we report the generation of a monospecific polyclonal antiserum which has helped to further elucidate the physical properties and developmental regulation of the cAMP receptor.

Original languageEnglish (US)
Pages (from-to)352-357
Number of pages6
JournalJournal of Biological Chemistry
Issue number1
StatePublished - 1987

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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