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The expression of a novel Ca2+/CaM stimulated adenylyl cyclase activity in the neuroblastoma cell line Lan-1 is regulated by cell density.

Abstract
In the human neuroblastoma cell line Lan-1, the mRNA encoding the Ca2+/calmodulin (CaM) sensitive adenylyl cyclase type-1 (AC-1) was detected by reverse transcription-polymerase chain reaction (RT-PCR) as well as by Northern blotting. However, neither Ca2+/CaM stimulated AC activity was found nor could AC-1 type protein be detected by a specific antibody (anti-1Cl). In contrast, when cells were grown to high cell density, Ca2+/CaM stimulated AC-activity could be indeed found in membranes. The large increase in activity was paralleled by the appearance of a 110 kDa protein detected by the monoclonal AC antibody BBC-2. At the same time a 150 kDa adenylyl cyclase species present in growing cells was absent. The 110 kDa protein co-migrated with bovine AC-1 and was slightly larger than the human AC-1. Unexpectedly, however, the antibody anti-1CI was not able to precipitate the newly induced Lan-1 AC. In addition, no increase in type-1 AC mRNA could be detected either by PCR or by Northern blotting. Treatment of Lan-1 cells with 10 microM retinoic acid for 7 days caused growth arrest and morphological differentiation of the cells, yet the induction of the Ca2+/CaM-stimulated AC activity was much lower than in the dense grown control cultures. It is concluded that the Ca2+/CaM-activated AC of M(r) 110 kDa in Lan-1 cells is not related to the previously known Ca2+/CaM stimulated AC isoforms, and might thus represent a novel AC.
AuthorsF W Kluxen, A Hülster, M Droste, T Pfeuffer
JournalBrain research (Brain Res) Vol. 740 Issue 1-2 Pg. 169-74 (Nov 18 1996) ISSN: 0006-8993 [Print] Netherlands
PMID8973811 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Calmodulin
  • Colforsin
  • Adenylyl Cyclases
  • Calcium
Topics
  • Adenylyl Cyclases (drug effects, metabolism)
  • Animals
  • Brain Neoplasms (metabolism)
  • Calcium (pharmacology)
  • Calmodulin (pharmacology)
  • Cattle
  • Cell Count
  • Colforsin (pharmacology)
  • Humans
  • Neuroblastoma (metabolism)
  • Tumor Cells, Cultured

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