Adenylyl cyclase type II is stimulated by PKC via C-terminal phosphorylation

Biochim Biophys Acta. 1997 Oct 11;1358(3):307-13. doi: 10.1016/s0167-4889(97)00073-6.


Adenylyl cyclases of the type II family differ from other subforms in that they are conditionally stimulated via alpha(s)/betagamma subunits and regulated by PKC mediated phosphorylation. AC II, stably expressed in HEK 239 cells, was incubated with the PKC activator tetradecanoylphorbol acetate (TPA). Using cells metabolically labeled with [32P]phosphate, TPA caused concerted stimulation of basal and forskolin activated adenylyl cyclase together with incorporation of [32P]phosphate into AC II protein. Enhanced phosphorylation was also indicated by a monoclonal anti-phosphothreonine antibody. Assignment of TPA-induced [32P]phosphate-incorporation to specific sites was achieved by a combination of chemical and immunochemical methods. Three out of five [32P]labeled peptides that were generated by fragmentation with N-chlorosuccinimide were also recognized by the monoclonal antibody BBC-4 [S. Mollner, T. Pfeuffer, Eur. J. Biochem. 171 (1988) 265-271] directed against an epitope 8 kDa from the extreme C-terminus. These findings suggested Ser-871 (consensus sequence ARSLK) and Thr-1057 (CTCR) as acceptor candidates of phorbolester induced phosphoryl transfer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Cell Line
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Isoenzymes / metabolism*
  • Isoquinolines / pharmacology
  • Kidney
  • Macromolecular Substances
  • Peptide Fragments / metabolism
  • Phosphates
  • Phosphorylation
  • Protein Kinase C / metabolism*
  • Protein Kinase Inhibitors
  • Protein Kinases / metabolism*
  • Recombinant Proteins / metabolism
  • Signal Transduction
  • Sulfonamides*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transfection


  • Enzyme Inhibitors
  • Isoenzymes
  • Isoquinolines
  • Macromolecular Substances
  • Peptide Fragments
  • Phosphates
  • Protein Kinase Inhibitors
  • Recombinant Proteins
  • Sulfonamides
  • Protein Kinases
  • Protein Kinase C
  • Adenylyl Cyclases
  • N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
  • Tetradecanoylphorbol Acetate