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. 1982 Oct 1;207(1):43–49. doi: 10.1042/bj2070043

Soluble adenylate cyclase activity in Neurospora crassa.

J A Reig, A R Kornblihtt, M M Flawiá, H N Torres
PMCID: PMC1153821  PMID: 6217814

Abstract

A soluble form of adenylate cyclase was extracted from mycelia of Neurospora crassa wild-type strains. This enzyme activity was purified by chromatography on hexyl-amino-Sepharose, agarose and Blue Sepharose and preparative polyacrylamide-gel electrophoresis. On sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, peak fractions from the later purification steps showed a main polypeptide band with an apparent molecular weight of about 66 000. The following hydrodynamic and molecular parameters were established for the Neurospora soluble adenylate cyclase activity: sedimentation coefficient, 6.25 S; Stokes radius, 7.3 nm; partial specific volume, 0.74 ml/g; molecular weight, 202 000; frictional ratio, 1.65. The isoelectric point of this enzyme activity was 4.65. The enzyme was not activated by GTP, [beta gamma-imido]GTP, fluoride or cholera toxin.

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Selected References

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  1. Braun T., Dods R. F. Development of a Mn-2+-sensitive, "soluble" adenylate cyclase in rat testis. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1097–1101. doi: 10.1073/pnas.72.3.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  3. Da Silveira J. F., Zingales B., Colli W. Characterization of an adenylyl cyclase activity in particulate preparations from epimastigote forms of Trypanosoma cruzi. Biochim Biophys Acta. 1977 Apr 12;481(2):722–733. doi: 10.1016/0005-2744(77)90306-0. [DOI] [PubMed] [Google Scholar]
  4. Flawiá M. M., Terenzi H. F., Torres H. N. Characterization of Neurospora crassa mutant stratins deficient in adenylate cyclase activity. Arch Biochem Biophys. 1977 Apr 30;180(2):334–342. doi: 10.1016/0003-9861(77)90046-7. [DOI] [PubMed] [Google Scholar]
  5. Flawiá M. M., Torres H. N. Adenylate cyclase activity in Neurospora crassa. I. General properties. J Biol Chem. 1972 Nov 10;247(21):6873–6879. [PubMed] [Google Scholar]
  6. Flawiá M. M., Torres H. N. Adenylate cyclase activity in Neurospora crassa. II. Kinetics. J Biol Chem. 1972 Nov 10;247(21):6880–6883. [PubMed] [Google Scholar]
  7. Gomes S. L., Mennucci L., da Costa Maia J. C. Adenylate cyclase activity and cyclic AMP metabolism during cytodifferentiation of Blastocladiella emersonii. Biochim Biophys Acta. 1978 Jun 15;541(2):190–198. doi: 10.1016/0304-4165(78)90392-6. [DOI] [PubMed] [Google Scholar]
  8. Kornblihtt A. R., Flawia M. M., Torres H. N. Manganese ion dependent adenylate cyclase activity in rat testes: purification and properties. Biochemistry. 1981 Mar 3;20(5):1262–1267. doi: 10.1021/bi00508a033. [DOI] [PubMed] [Google Scholar]
  9. Neer E. J. Physical and functional properties of adenylate cyclase from mature rat testis. J Biol Chem. 1978 Aug 25;253(16):5808–5812. [PubMed] [Google Scholar]
  10. Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
  11. Paveto C., Epstein A., Passeron S. Studies on cyclic adenosine 3' ,5'-monophosphate levels, Adenylate cyclase and phosphodiesterase activities in the dimorphic fungus Mucor rouxii. Arch Biochem Biophys. 1975 Aug;169(2):449–457. doi: 10.1016/0003-9861(75)90187-3. [DOI] [PubMed] [Google Scholar]
  12. Ross E. M., Howlett A. C., Ferguson K. M., Gilman A. G. Reconstitution of hormone-sensitive adenylate cyclase activity with resolved components of the enzyme. J Biol Chem. 1978 Sep 25;253(18):6401–6412. [PubMed] [Google Scholar]
  13. Terenzi H. F., Flawia M. M., Tellez-Inon M. T., Torres H. N. Control of Neurospora crassa morphology by cyclic adenosine 3', 5'-monophosphate and dibutyryl cyclic adenosine 3', 5'-monophosphate. J Bacteriol. 1976 Apr;126(1):91–99. doi: 10.1128/jb.126.1.91-99.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Varimo K., Londesborough J. Solubilization and other studies on adenylate cyclase of baker's yeast. Biochem J. 1976 Nov;159(2):363–370. doi: 10.1042/bj1590363. [DOI] [PMC free article] [PubMed] [Google Scholar]

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