PMID: 2469Jan 15, 1976

Guanylate cyclase: assay and properties of the particulate and supernatant enzymes in mouse parotid

European Journal of Biochemistry
J P Durham

Abstract

A new, very sensitive, rapid and reliable assay for guanylate cyclase has been established based on conversion of [32P]GTP to [32P]guanosine 3':5'-monophosphate and its separation on Dowex 50 and aluminium oxide columns. The optimum conditions for the assay of mouse parotid guanylate cyclase have been established and using this procedure the properties of the enzyme have been investigated. The enzyme was found in both the particulate and supernatant fractions. The particulate enzyme was activated 12-fold by Triton X-100 and the supernatant enzyme activity increased 2-fold. In the presence of detergent guanylate cyclase activity was distributed 85% in the particulate and 15% in the supernatant fractions, respectively. The particulate activity was localised in a plasma membrane fraction. Guanylate cyclase activity was also assayed in a wide variety of other tissues. In all cases enzymatic activity was found in both the particulate and supernatant fractions. The distribution varied with the tissue but only the intestinal mucosa had a greater proportion of total guanylate cyclase activity in the particulate fraction than the parotid. The two enzymes showed some similar properties. Their pH optima were pH 7.4, both enzymes were inhi...Continue Reading

References

Jan 23, 1975·Biochimica Et Biophysica Acta·L E Limbird, R J Lefkowitz
Apr 12, 1973·Biochimica Et Biophysica Acta·W Joseph ThompsonS A Little
Nov 1, 1973·Archives of Biochemistry and Biophysics·W Joseph ThompsonS A Little
Aug 1, 1974·The Biochemical Journal·S L Howell, W Montague
Jan 1, 1969·Biochimica Et Biophysica Acta·A A White, G D Aurbach
Dec 30, 1971·Annals of the New York Academy of Sciences·J G HardmanE W Sutherland
Oct 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·J W HaddenN D Goldberg
Jun 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·R Kram, G M Tomkins
Dec 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·G SchultzE W Sutherland
Aug 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·G IllianoP Cuatrecasas
Oct 1, 1972·Analytical Biochemistry·P J Geiger, S P Bessman
Jun 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·W J GeorgeN D Goldberg
Jan 1, 1964·Progress in Nucleic Acid Research and Molecular Biology·P Mandel

Citations

Apr 28, 2009·Biochimie·Priyaranjan PattanaikFocco van den Akker
Oct 1, 2005·Current Protocols in Pharmacology·John J KellyRoland Seifert

Related Concepts

Aluminum
Chromatography
Ion-Exchange Chromatography Procedure
Enzyme Activation
Guanylate Cyclase
Hydrogen-Ion Concentration
Hydroxymercuribenzoates
Alloenzymes
Manganese
Tissue Specificity

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