NDLI: Identification of the catalytic subunit of brain adenylate cyclase: a calmodulin binding protein of 135 kDa.

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Identification of the catalytic subunit of brain adenylate cyclase: a calmodulin binding protein of 135 kDa.

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Identification of the catalytic subunit of brain adenylate cyclase: a calmodulin binding protein of 135 kDa.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	D'alayer, J. Haiech, J. Coussen, F. Monneron, A.
Abstract	The partial purification of the eukaryote adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] catalytic subunit has been achieved by a procedure based on the calmodulin (CaM) sensitivity of the enzyme. Small amounts of rat brain synaptosomal membranes depleted of CaM were solubilized with Lubrol and subjected to a three-step chromatographic procedure involving gel filtration, a CaM-Sepharose affinity step, and fast protein liquid chromatography. About 20% of the adenylate cyclase activity contained in the membranes was recovered in the final enriched fraction with a specific activity of 200 nmol X mg-1 X min-1. The alpha subunits of the adenylate cyclase stimulatory proteins NS were absent from this final fraction. The addition of CaM, of forskolin, or of preactivated NS-containing fractions to this preparation greatly increased the enzyme activity. A CaM-binding polypeptide of 135,000 Da copurified with the adenylate cyclase activity in each of the three steps. Polyacrylamide gel electrophoresis of the final fraction showed that this polypeptide represented 35% of the total protein. We propose that this polypeptide is likely to be the adenylate cyclase catalytic subunit. This enzyme would represent close to 0.5% of the synaptosomal membrane proteins. Its low turnover number would be due to the absence of the alpha subunits of the NS regulatory proteins and would correspond to the enzymic basal level.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	20
Volume Number	82
Language	English
Publisher	National Academy of Sciences
Publisher Date	1985-11-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Adenylate Cyclase Isolation & Purification Brain Enzymology Calmodulin-Binding Proteins Synaptosomes Metabolism Animals Intracellular Membranes Kinetics Molecular Weight Research Support, Non-U.S. Gov't Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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