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PMID: 6933510 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Partial purification and characterization of a macromolecule which enhances fluoride activation of adenylate cyclase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 77 ·No. 8 ·1980-08-00 ·Pages 4628-32

Rasenick MM, Bitensky MW

Abstract

Fluoride activation of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] is significantly enhanced (2 to 5 times) by a protein factor isolated from rat brain. The fluoride-dependent adenylate cyclase stimulator (FCS) is nondialyzable, trypsin-labile, and stable at 90 degrees C for 10 min. FCS stimulates adenylate cyclase activity only in the presence of NaF (2-25 mM) and this effect is independent of added GTP, 5'-guanylylimidodiphosphate, or calcium. FCS has been purified roughly 3000-fold from a 12,000 X g supernatant fraction of rat brain homogenate. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis and sucrose density gradient sedimentation suggest that FCS is a monomer with an apparent Mr of 59,000. Isoelectric focusing indicates FCS has a pI of 8.9. FCS from rat brain stimulates fluoride-activated adenylate cyclase from a variety of cell types, and FCS can also be isolated from rat liver. The effects of FCS are not reversed by washing membranes when the membranes and FCS are preincubated with NaF. The Km of adenylate cyclase for ATP and the fluoride concentration causing half-maximal activation are unchanged by FCS; however, FCS increases the Vmax by 2.5-fold. FCS may act to increase the catalytic efficiency of fluoride-activated complexes of the GTP-binding unit with adenylate cyclase or to enhance the formation of additional active complexes.

MeSH Terms
Adenylyl Cyclases/metabolism Animals Brain/enzymology Calcium/metabolism Enzyme Activation/drug effects Fluorides/pharmacology Isoelectric Point Kinetics Liver/enzymology Male Molecular Weight Rats Synaptic Membranes/enzymology
Chemicals
Adenylyl Cyclases Fluorides Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rasenick M M
Bitensky M W
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33 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.77.8.4628
Published
1980-08-00
Pages
4628-32
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC349898
Subset
IM
Grants
NIADDK NIH HHS · 1 F32 AM 06278 · United States
NIGMS NIH HHS · 5T32 GM 07562 · United States
NIADDK NIH HHS · R01 AM 20179 · United States
External Links
PubMed source DOI: 10.1073/pnas.77.8.4628 PMC Full Text View journal details
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