A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis

T. D. Sutherland, G. C. Unnithan, J. F. Andersen, P. H. Evans, M. B. Murataliev, L. Z. Szabo, Eugene A Mash, W. S. Bowers, R. Feyereisen

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine glands that produce the insect juvenile hormone (JH) is most closely related to P450 proteins of family 4 and was named CYP4C7. The CYP4C7 gene is expressed selectively in the CA; its message could not be detected in the fat body, corpora cardiaca, or brain, but trace levels of expression were found in the midgut and caeca. The levels of CYP4C7 mRNA in the CA, measured by ribonuclease protection assays, were linked to the activity cycle of the glands. In adult females, CYP4C7 expression increased immediately after the peak of JH synthesis, reaching a maximum on day 7, just before oviposition. mRNA levels then declined after oviposition and during pregnancy. The CYP4C7 protein was produced in Escherichia coli as a C-terminal His-tagged recombinant protein. In a reconstituted system with insect NADPH cytochrome P450 reductase, cytochrome b(s), and NADPH, the purified CYP4C7 metabolized (2E,6E)-farnesol to a more polar product that was identified by GC-MS and by NMR as (10E)-12- hydroxyfarnesol, CYP4C7 converted JH III to 12-trans-hydroxy JH HI and metabolized other JH-like sesquiterpenoids as well. This ω-hydroxylation of sesquiterpenoids appears to be a metabolic pathway in the corpora allata that may play a role in the suppression of JH biosynthesis at the end of the gonotrophic cycle.

Original languageEnglish (US)
Pages (from-to)12884-12889
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number22
DOIs
StatePublished - Oct 27 1998

Fingerprint

Insect Hormones
Corpora Allata
Juvenile Hormones
Terpenes
Mixed Function Oxygenases
Cytochrome P-450 Enzyme System
Oviposition
Farnesol
Endocrine Glands
Activity Cycles
NADPH-Ferrihemoprotein Reductase
Messenger RNA
Cockroaches
Cytochromes b
Fat Body
Hydroxylation
Ribonucleases
Metabolic Networks and Pathways
NADP
Gene Library

Keywords

  • Corpora allata
  • CYP4C7
  • NMR
  • P450 reconstitution
  • Sesquiterpenoid

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Sutherland, T. D., Unnithan, G. C., Andersen, J. F., Evans, P. H., Murataliev, M. B., Szabo, L. Z., ... Feyereisen, R. (1998). A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis. Proceedings of the National Academy of Sciences of the United States of America, 95(22), 12884-12889. https://doi.org/10.1073/pnas.95.22.12884

A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis. / Sutherland, T. D.; Unnithan, G. C.; Andersen, J. F.; Evans, P. H.; Murataliev, M. B.; Szabo, L. Z.; Mash, Eugene A; Bowers, W. S.; Feyereisen, R.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, No. 22, 27.10.1998, p. 12884-12889.

Research output: Contribution to journalArticle

Sutherland, TD, Unnithan, GC, Andersen, JF, Evans, PH, Murataliev, MB, Szabo, LZ, Mash, EA, Bowers, WS & Feyereisen, R 1998, 'A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis', Proceedings of the National Academy of Sciences of the United States of America, vol. 95, no. 22, pp. 12884-12889. https://doi.org/10.1073/pnas.95.22.12884
Sutherland, T. D. ; Unnithan, G. C. ; Andersen, J. F. ; Evans, P. H. ; Murataliev, M. B. ; Szabo, L. Z. ; Mash, Eugene A ; Bowers, W. S. ; Feyereisen, R. / A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis. In: Proceedings of the National Academy of Sciences of the United States of America. 1998 ; Vol. 95, No. 22. pp. 12884-12889.
@article{fa125a5a33cd4b46a3d4f051d8d0c757,
title = "A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis",
abstract = "A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine glands that produce the insect juvenile hormone (JH) is most closely related to P450 proteins of family 4 and was named CYP4C7. The CYP4C7 gene is expressed selectively in the CA; its message could not be detected in the fat body, corpora cardiaca, or brain, but trace levels of expression were found in the midgut and caeca. The levels of CYP4C7 mRNA in the CA, measured by ribonuclease protection assays, were linked to the activity cycle of the glands. In adult females, CYP4C7 expression increased immediately after the peak of JH synthesis, reaching a maximum on day 7, just before oviposition. mRNA levels then declined after oviposition and during pregnancy. The CYP4C7 protein was produced in Escherichia coli as a C-terminal His-tagged recombinant protein. In a reconstituted system with insect NADPH cytochrome P450 reductase, cytochrome b(s), and NADPH, the purified CYP4C7 metabolized (2E,6E)-farnesol to a more polar product that was identified by GC-MS and by NMR as (10E)-12- hydroxyfarnesol, CYP4C7 converted JH III to 12-trans-hydroxy JH HI and metabolized other JH-like sesquiterpenoids as well. This ω-hydroxylation of sesquiterpenoids appears to be a metabolic pathway in the corpora allata that may play a role in the suppression of JH biosynthesis at the end of the gonotrophic cycle.",
keywords = "Corpora allata, CYP4C7, NMR, P450 reconstitution, Sesquiterpenoid",
author = "Sutherland, {T. D.} and Unnithan, {G. C.} and Andersen, {J. F.} and Evans, {P. H.} and Murataliev, {M. B.} and Szabo, {L. Z.} and Mash, {Eugene A} and Bowers, {W. S.} and R. Feyereisen",
year = "1998",
month = "10",
day = "27",
doi = "10.1073/pnas.95.22.12884",
language = "English (US)",
volume = "95",
pages = "12884--12889",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "22",

}

TY - JOUR

T1 - A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis

AU - Sutherland, T. D.

AU - Unnithan, G. C.

AU - Andersen, J. F.

AU - Evans, P. H.

AU - Murataliev, M. B.

AU - Szabo, L. Z.

AU - Mash, Eugene A

AU - Bowers, W. S.

AU - Feyereisen, R.

PY - 1998/10/27

Y1 - 1998/10/27

N2 - A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine glands that produce the insect juvenile hormone (JH) is most closely related to P450 proteins of family 4 and was named CYP4C7. The CYP4C7 gene is expressed selectively in the CA; its message could not be detected in the fat body, corpora cardiaca, or brain, but trace levels of expression were found in the midgut and caeca. The levels of CYP4C7 mRNA in the CA, measured by ribonuclease protection assays, were linked to the activity cycle of the glands. In adult females, CYP4C7 expression increased immediately after the peak of JH synthesis, reaching a maximum on day 7, just before oviposition. mRNA levels then declined after oviposition and during pregnancy. The CYP4C7 protein was produced in Escherichia coli as a C-terminal His-tagged recombinant protein. In a reconstituted system with insect NADPH cytochrome P450 reductase, cytochrome b(s), and NADPH, the purified CYP4C7 metabolized (2E,6E)-farnesol to a more polar product that was identified by GC-MS and by NMR as (10E)-12- hydroxyfarnesol, CYP4C7 converted JH III to 12-trans-hydroxy JH HI and metabolized other JH-like sesquiterpenoids as well. This ω-hydroxylation of sesquiterpenoids appears to be a metabolic pathway in the corpora allata that may play a role in the suppression of JH biosynthesis at the end of the gonotrophic cycle.

AB - A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine glands that produce the insect juvenile hormone (JH) is most closely related to P450 proteins of family 4 and was named CYP4C7. The CYP4C7 gene is expressed selectively in the CA; its message could not be detected in the fat body, corpora cardiaca, or brain, but trace levels of expression were found in the midgut and caeca. The levels of CYP4C7 mRNA in the CA, measured by ribonuclease protection assays, were linked to the activity cycle of the glands. In adult females, CYP4C7 expression increased immediately after the peak of JH synthesis, reaching a maximum on day 7, just before oviposition. mRNA levels then declined after oviposition and during pregnancy. The CYP4C7 protein was produced in Escherichia coli as a C-terminal His-tagged recombinant protein. In a reconstituted system with insect NADPH cytochrome P450 reductase, cytochrome b(s), and NADPH, the purified CYP4C7 metabolized (2E,6E)-farnesol to a more polar product that was identified by GC-MS and by NMR as (10E)-12- hydroxyfarnesol, CYP4C7 converted JH III to 12-trans-hydroxy JH HI and metabolized other JH-like sesquiterpenoids as well. This ω-hydroxylation of sesquiterpenoids appears to be a metabolic pathway in the corpora allata that may play a role in the suppression of JH biosynthesis at the end of the gonotrophic cycle.

KW - Corpora allata

KW - CYP4C7

KW - NMR

KW - P450 reconstitution

KW - Sesquiterpenoid

UR - http://www.scopus.com/inward/record.url?scp=0032573065&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032573065&partnerID=8YFLogxK

U2 - 10.1073/pnas.95.22.12884

DO - 10.1073/pnas.95.22.12884

M3 - Article

VL - 95

SP - 12884

EP - 12889

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 22

ER -