A Brain Synaptosomal Adenylyl Cyclase of High Specific Activity Is Photolabeled with Azido-ATP

F. Castets, G. Baillat, S. Mirzoeva, K. Mabrouk, J. Garin, J. D'Alayer, A. Monneron

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Partially purified adenylyl cyclase preparations of high specific activity (60 ± 10 µmol cAMP/(mg·min)) were obtained from rat brain synaptosomes (Orlando, C., d'Alayer, J., Baillat, G., Castets, F., Jeannequin, O., Mazié, J. C., & Monneron, A. (1992) Biochemistry 31, 3215–3222). Adenylyl cyclase activity was stimulated 4-fold by Ca2+/calmodulin and 2-fold by forskolin or by Mn2+. These preparations contained two major proteins of 140 and 110 kDa. The 140-kDa protein was identified as the neural cell adhesion molecule. The 11 ?-kDa protein was specifically recognized by affinity-purified antibodies directed against a peptide corresponding to sequence 976–1013 of adenylyl cyclase type I. It was photolabeled by [α-32P]8- and 2-N3ATP in a light-dependent manner and was by far the most heavily labeled component of FC fractions. Saturation was obtained with 30 µ? [32P]8-N3ATP. Photoinsertion of N3ATP into the protein was largely prevented by ATP or adenylyl imidodiphosphate but not by ADP, AMP, or adenosine. A modest incorporation of N3cAMP and photoinsertion of [α-32P]N3GTP into the 110-kDa protein were observed. Although some of the properties of the synaptosomal 110-kDa protein described here would match those expected from adenylyl cyclase type I, it appears that its specific activity is on the order of 1 mmol cAMP/(mg·min), about 200-fold that measured for brain adenylyl cyclases type I.

Original languageEnglish (US)
Pages (from-to)5063-5069
Number of pages7
JournalBiochemistry
Volume33
Issue number17
DOIs
StatePublished - May 1 1994

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'A Brain Synaptosomal Adenylyl Cyclase of High Specific Activity Is Photolabeled with Azido-ATP'. Together they form a unique fingerprint.

  • Cite this

    Castets, F., Baillat, G., Mirzoeva, S., Mabrouk, K., Garin, J., D'Alayer, J., & Monneron, A. (1994). A Brain Synaptosomal Adenylyl Cyclase of High Specific Activity Is Photolabeled with Azido-ATP. Biochemistry, 33(17), 5063-5069. https://doi.org/10.1021/bi00183a009