PMID: 6108559Oct 1, 1980Paper

Products of reaction catalyzed by purified rat liver guanylate cyclase determined by 31p NMR spectroscopy

Proceedings of the National Academy of Sciences of the United States of America
S C TsaiM Vaughan

Abstract

Products of the reactions catalyzed by highly purified preparations of soluble guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] from rat liver were identified and quantified with 31P NMR spectroscopy. Utilization of this technique necessitated modification of the standard assay conditions; higher concentrations of enzyme and substrate (2 mM), Mg2+ instead of Mn2+, and longer incubation times (up to 46 hr) at 30 degrees C were used. Revision of our reported procedure for purificaton of guanylate cyclase [Tsai, S-C., Manganiello, V. C. & Vaughan, M. (1978) J. Biol. Chem. 253, 8452-8457] to include chromatography on Ultrogel AcA34 and agarose-hexane-GTP provided an enzyme with specific activity higher than in our earlier preparations. 31P NMR spectra obtained during incubation of this enzyme showed that the rates of GTP disappearance and cyclic GMP (cGMP) accumulation were constant for approximately 16 hr. They indicated, however, that the preparations were contaminated with inorganic pyrophosphatase. This was removed by preparative electrophoresis, yielding enzyme with specific activities (900-1300 nmol/min per mg of protein) higher than those reported for guanylate cyclases from rat liver or lung. With this pr...Continue Reading

References

Jan 1, 1975·Archives of Biochemistry and Biophysics·D L GarbersJ G Hardman

❮ Previous
Next ❯

Citations

May 29, 1981·Biochemical and Biophysical Research Communications·S C TsaiM Vaughan

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biosynthetic Transformations

Biosyntheic transformtions are multi-step, enzyme-catalyzed processes where substrates are converted into more complex products in living organisms. Simple compounds are modified, converted into other compounds, or joined together to form macromolecules. Discover the latest research on biosynthetic transformations here.

Related Papers

Proceedings of the National Academy of Sciences of the United States of America
J M BraughlerF Murad
Proceedings of the National Academy of Sciences of the United States of America
W T ShierN M Thanassi
© 2021 Meta ULC. All rights reserved