Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1978 Aug;75(8):3693–3697. doi: 10.1073/pnas.75.8.3693

Properties of the interaction of fluoride- and guanylyl-5'-imidodiphosphate-regulatory proteins with adenylate cyclase.

M Hebdon, H Le Vine 3rd, N Sahyoun, C J Schmitges, P Cuatrecasas
PMCID: PMC392852  PMID: 278984

Abstract

The mechanism of activation of adenylate cyclase by guanylyl-5'-imidodiphosphate [Gpp(NH)p] and NaF has been investigated by studying the reconstitution of Gpp(NH)p and NaF sensitivity of an enzyme rendered insensitive to these agents by differential detergent extraction of a particulate brain enzyme. Such reconstitution can be achieved by the addition of macromolecular regulatory factors from membranes of various tissues. Trypsin digestion and thermal inactivation provide evidence for the existence of two distinct regulatory functions, one capable of restoring the Gpp(NH)p response and another the NaF response. The regulatory protein(s) seem to interact with their respective activators in an easily reversible, divalent cation-independent reaction. This appears to be followed by a high-affinity interaction between the catalytic and regulatory components of adenylate cyclase in a slow, temperature-dependent, divalent cation-dependent process tha produces the persistently activated state of the enzyme. The enzyme activation can be reversed by methods that separate catalytic from regulatory components and the resulting enzyme activity can be restimulated by the reconstitution technique.

Full text

PDF
3693

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bennett V., Cuatrecasas P. Mechanism of action of Vibrio cholerae enterotoxin. Effects on adenylate cyclase of toad and rat erythrocyte plasma membranes. J Membr Biol. 1975 Jun 3;22(1):1–28. doi: 10.1007/BF01868161. [DOI] [PubMed] [Google Scholar]
  2. Bradham L. S. Fluoride activation of rat brain adenylate cyclase: the requirement for a protein co-factor. J Cyclic Nucleotide Res. 1977 Apr;3(2):119–128. [PubMed] [Google Scholar]
  3. Cassel D., Selinger Z. Catecholamine-induced release of [3H]-Gpp(NH)p from turkey erythrocyte adenylate cyclase. J Cyclic Nucleotide Res. 1977 Feb;3(1):11–22. [PubMed] [Google Scholar]
  4. Cassel D., Selinger Z. Mechanism of adenylate cyclase activation by cholera toxin: inhibition of GTP hydrolysis at the regulatory site. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3307–3311. doi: 10.1073/pnas.74.8.3307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cuatrecasas P., Jacobs S., Bennett V. Activation of adenylate cyclase by phosphoramidate and phosphonate analogs of GTP: possible role of covalent enzyme-substrate intermediates in the mechanism of hormonal activation. Proc Natl Acad Sci U S A. 1975 May;72(5):1739–1743. doi: 10.1073/pnas.72.5.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jacobs S., Bennett V., Cuatrecasas P. Kinetics of irreversible activation of adenylate cyclase of fat cell membranes by phosphonium and phosphoramidate analogs of gtp1. J Cyclic Nucleotide Res. 1976 Jul-Aug;2(4):205–223. [PubMed] [Google Scholar]
  7. Lynch T. J., Tallant E. A., Cheung W. Y. Rat brain adenylate cyclase. Further studies on its stimulation by a Ca2+-binding protein. Arch Biochem Biophys. 1977 Jul;182(1):124–133. doi: 10.1016/0003-9861(77)90290-9. [DOI] [PubMed] [Google Scholar]
  8. Pfeuffer T. GTP-binding proteins in membranes and the control of adenylate cyclase activity. J Biol Chem. 1977 Oct 25;252(20):7224–7234. [PubMed] [Google Scholar]
  9. Ross E. M., Gilman A. G. Reconstitution of catecholamine-sensitive adenylate cyclase activity: interactions of solubilized components with receptor-replete membranes. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3715–3719. doi: 10.1073/pnas.74.9.3715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ross E. M., Gilman A. G. Resolution of some components of adenylate cyclase necessary for catalytic activity. J Biol Chem. 1977 Oct 25;252(20):6966–6969. [PubMed] [Google Scholar]
  11. Sahyoun N., Schmitges C. J., Le Vine H., 3rd, Cuatrecasas P. Molecular resolution and reconstitution of the GPP (NH) P and NAF sensitive adenylate cyclase system. Life Sci. 1977 Dec 15;21(12):1857–1863. doi: 10.1016/0024-3205(77)90169-2. [DOI] [PubMed] [Google Scholar]
  12. Sevilla N., Levitzki A. The activation of adenylate cyclase by 1-epinephrine and guanylylimidodiphosphate and its reversal by 1-epinephrine and GTP. FEBS Lett. 1977 Apr 1;76(1):129–134. doi: 10.1016/0014-5793(77)80136-1. [DOI] [PubMed] [Google Scholar]
  13. Wheeler G. L., Bitensky M. W. A light-activated GTPase in vertebrate photoreceptors: regulation of light-activated cyclic GMP phosphodiesterase. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4238–4242. doi: 10.1073/pnas.74.10.4238. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES