NDLI: Choleragen activation of solubilized adenylate cyclase: requirement for GTP and protein activator for demonstration of enzymatic activity.

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Choleragen activation of solubilized adenylate cyclase: requirement for GTP and protein activator for demonstration of enzymatic activity.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Vaughan, M. Moss, J.
Abstract	The requirements for choleragen activation of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] were investigated by using an enzyme preparation solubilized with Triton X-100 from an extensively washed brain particulate fraction and partially purified with DEAE-cellulose. Unlike the particulate enzyme, this preparation was not activated after incubation with choleragen plus dithiothreitol, ATP, and NAD. Addition of the purified protein activator of cyclic nucleotide phosphodiesterase and calcium to the partially purified enzyme increased basal activity somewhat, but choleragen activation was minimal. When cyclase was incubated with GTP plus the protein activator (and calcium), choleragen markedly increased the activity 3- to 6-fold. When GppNHp and protein activator were incubated with the cyclase prior to assay, activity was elevated but no effect of choleragen was observed. GTP and GppNHp had relatively small effects on cyclase activity in the absence of protein activator or if they were added directly to the assay. Boiled brain supernatant was consistently more effective than protein activator (plus calcium) and GTP, suggesting that other factors are required for maximal cyclase activity after choleragen treatment. It appears that the cyclase system is dissociable into several components, all of which may be necessary for optimal regulation of activity. It is probable that one of these is the heat-stable calcium-dependent protein activator of cyclic nucleotide phosphodiesterase and adenylate cyclase that we have found is required along with GTP for demonstration of choleragen activation of partially purified brain adenylate cyclase.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	10
Volume Number	74
Language	English
Publisher	National Academy of Sciences
Publisher Date	1977-01-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Adenylate Cyclase Metabolism Brain Enzymology Cholera Toxin Pharmacology Guanosine Triphosphate Nerve Tissue Proteins Adenosine Triphosphate Animals Calcium Egtazic Acid Enzyme Activation Drug Effects Guanylyl Imidodiphosphate In Vitro Techniques NAD Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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