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PMID: 230476 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Binding of inhibitor alters kinetic and physical properties of extracellular cyclic AMP phosphodiesterase from Dictyostelium discoideum.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 76 ·No. 11 ·1979-11-00 ·Pages 5450-4

Kessin RH, Orlow SJ, Shapiro RI, Franke J

Abstract

The extracellular adenosine 3',5'-cyclic monophosphate phosphodiesterase (3',5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 3.1.4.17) produced by Dictyostelium discoideum has two kinetic forms. The free enzyme has a Km of approximately 10 microM. The second form is the result of a complex formed with a heat-stable inhibitor and has a Km in the millimolar range. Treating the enzyme-inhibitor complex with dithiothreitol stimulated enzyme activity 20- to 100-fold and changed in Km from millimolar to micromolar. Dithiothreitol inactivated the inhibitor. Reconstituting purified enzyme with excess inhibitor returned the Km to the millimolar range. Under conditions known to inhibit the production of extracellular inhibitor or in mutants that lack it, extracellular phosphodiesterase activity was already high and could not be increased by dithiothreitol. The phosphodiesterase and inhibitor sedimented at 6 S and 3 S, respectively; the enzyme-inhibitor complex sedimented at 6.7 S.

MeSH Terms
3',5'-Cyclic-AMP Phosphodiesterases/antagonists & inhibitors Dictyostelium/enzymology Dithiothreitol/pharmacology Enzyme Activation/drug effects Extracellular Space/enzymology Hot Temperature Kinetics Molecular Weight
Chemicals
3',5'-Cyclic-AMP Phosphodiesterases Dithiothreitol
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kessin R H
Orlow S J
Shapiro R I
Franke J
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23 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.76.11.5450
Published
1979-11-00
Pages
5450-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC411666
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.76.11.5450 PMC Full Text View journal details
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