PMID: 488526Sep 1, 1979Paper

Ecdysone 20-monooxygenase: characterization of an insect cytochrome p-450 dependent steroid hydroxylase

Molecular and Cellular Endocrinology
S L SmithL I Gilbert

Abstract

Ecdysone 20-monooxygenase, the enzyme system that hydroxylates ecdysone at C-20 of the side-chain to form ecdysterone, has been characterized in the fat body of early last instar larvae of the tobacco hornworm, Manduca sexta, using a radioenzymological assay. Ecdysterone was demonstrated to be the product of the enzyme system by high-pressure liquid chromatography, gas-liquid chromatography and mass spectrometry. Differential centrifugation, sucrose-gradient centrifugation, electron microscopy and organelle-marker enzyme analysis revealed that ecdysone 20-monooxygenase activity is associated with the mitochondria. The enzymatic properties of ecdysone 20-monooxygenase are that it is most active in a 0.05 M phosphate buffer, is inhibited by Mg2+ and exhibits pH and temperature optima at 7.5 and 30 degrees C, respectively. The enzyme complex has an apparent Km for ecdysone of 1.60 x 10(-7) M and is competitively inhibited by its product, ecdysterone, with an apparent Ki of 2.72 x 10(-5) M. The cytochrome P-450 nature of this insect steroid hydroxylase was initially suggested by its obligate requirement for NADPH and its inhibition by carbon monoxide, p-chloromercuribenzoate, metyrapone and p-aminoglutethimide but not by cyanide. D...Continue Reading

References

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Citations

Jul 7, 2011·The Journal of Membrane Biology·Kurt P VandockAlyssa A Rassman
Aug 19, 2007·Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology·Inga Yu RauschenbachA A Alekseev
Apr 27, 2000·Lipids·Y FujimotoM Morisaki
Aug 1, 1983·Molecular and Cellular Endocrinology·S L SmithL I Gilbert
Jan 1, 1993·Insect Biochemistry and Molecular Biology·M L GrieneisenL I Gilbert
Jul 31, 2001·Molecular and Cellular Endocrinology·J T WarrenL I Gilbert
May 20, 2005·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Thomas L Poulos
Nov 5, 2003·Proceedings of the National Academy of Sciences of the United States of America·Thomas L Poulos
Nov 12, 2003·Proceedings of the National Academy of Sciences of the United States of America·Anna PetrykMichael B O'Connor
May 11, 2010·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Christopher A DrummondCarmen F Fioravanti
Oct 8, 2008·Archives of Insect Biochemistry and Physiology·Kurt P VandockCarmen F Fioravanti
Apr 24, 2012·Archives of Insect Biochemistry and Physiology·Kurt P VandockCarmen F Fioravanti
Feb 14, 2006·Molecular and Cellular Endocrinology·Kim F RewitzLawrence I Gilbert
Oct 1, 1989·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·M J Ronis, E Hodgson
Apr 25, 2000·Archives of Biochemistry and Biophysics·D R WilliamsH H Rees
Jul 29, 1988·Biochemical and Biophysical Research Communications·S L Smith, M J Mitchell
Nov 13, 1987·Biochemical and Biophysical Research Communications·J SrivatsanM Agosin

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