Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta

Mark Wogulis, Craig E. Wheelock, Shizuo G. Kamita, Andrew C. Hinton, Paul A. Whetstone, Bruce D. Hammock, David K. Wilson

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Juvenile hormone (JH) is an insect hormone containing an α/β-unsaturated ester consisting of a small alcohol and long, hydrophobic acid. JH degradation is required for proper insect development. One pathway of this degradation is through juvenile hormone esterase (JHE), which cleaves the JH ester bond to produce methanol and JH acid. JHE is a member of the functionally divergent α/β-hydrolase family of enzymes and is a highly efficient enzyme that cleaves JH at very low in vivo concentrations. We present here a 2.7 Å crystal structure of JHE from the tobacco hornworm Manduca sexta (MsJHE) in complex with the transition state analogue inhibitor 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP) covalently bound to the active site. This crystal structure, the first JHE structure reported, contains a long, hydrophobic binding pocket with the solvent-inaccessible catalytic triad located at the end. The structure explains many of the interactions observed between JHE and its substrates and inhibitors, such as the preference for small alcohol groups and long hydrophobic backbones. The most potent JHE inhibitors identified to date contain a trifluoromethyl ketone (TFK) moiety and have a sulfur atom α to the ketone. In this study, sulfur-aromatic interactions were observed between the sulfur atom of OTFP and a conserved aromatic residue in the crystal structure. Mutational analysis supported the hypothesis that these interactions contribute to the potency of sulfur-containing TFK inhibitors. Together, these results clarify the binding mechanism of JHE inhibitors and provide useful observations for the development of additional enzyme inhibitors for a variety of enzymes.

Original languageEnglish (US)
Pages (from-to)4045-4057
Number of pages13
JournalBiochemistry
Volume45
Issue number13
DOIs
StatePublished - Apr 4 2006

Fingerprint

Manduca
Juvenile Hormones
Insects
Sulfur
Ketones
Crystal structure
Esters
Enzymes
Insect Hormones
Alcohols
Degradation
Atoms
Acids
Tobacco
Enzyme Inhibitors
Hydrolases
juvenile hormone esterase
Methanol
Catalytic Domain
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Wogulis, M., Wheelock, C. E., Kamita, S. G., Hinton, A. C., Whetstone, P. A., Hammock, B. D., & Wilson, D. K. (2006). Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta. Biochemistry, 45(13), 4045-4057. https://doi.org/10.1021/bi0521644

Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta. / Wogulis, Mark; Wheelock, Craig E.; Kamita, Shizuo G.; Hinton, Andrew C.; Whetstone, Paul A.; Hammock, Bruce D.; Wilson, David K.

In: Biochemistry, Vol. 45, No. 13, 04.04.2006, p. 4045-4057.

Research output: Contribution to journalArticle

Wogulis, M, Wheelock, CE, Kamita, SG, Hinton, AC, Whetstone, PA, Hammock, BD & Wilson, DK 2006, 'Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta', Biochemistry, vol. 45, no. 13, pp. 4045-4057. https://doi.org/10.1021/bi0521644
Wogulis M, Wheelock CE, Kamita SG, Hinton AC, Whetstone PA, Hammock BD et al. Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta. Biochemistry. 2006 Apr 4;45(13):4045-4057. https://doi.org/10.1021/bi0521644
Wogulis, Mark ; Wheelock, Craig E. ; Kamita, Shizuo G. ; Hinton, Andrew C. ; Whetstone, Paul A. ; Hammock, Bruce D. ; Wilson, David K. / Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta. In: Biochemistry. 2006 ; Vol. 45, No. 13. pp. 4045-4057.
@article{938dba8cc9284715bc9a2c21423eb32f,
title = "Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta",
abstract = "Juvenile hormone (JH) is an insect hormone containing an α/β-unsaturated ester consisting of a small alcohol and long, hydrophobic acid. JH degradation is required for proper insect development. One pathway of this degradation is through juvenile hormone esterase (JHE), which cleaves the JH ester bond to produce methanol and JH acid. JHE is a member of the functionally divergent α/β-hydrolase family of enzymes and is a highly efficient enzyme that cleaves JH at very low in vivo concentrations. We present here a 2.7 {\AA} crystal structure of JHE from the tobacco hornworm Manduca sexta (MsJHE) in complex with the transition state analogue inhibitor 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP) covalently bound to the active site. This crystal structure, the first JHE structure reported, contains a long, hydrophobic binding pocket with the solvent-inaccessible catalytic triad located at the end. The structure explains many of the interactions observed between JHE and its substrates and inhibitors, such as the preference for small alcohol groups and long hydrophobic backbones. The most potent JHE inhibitors identified to date contain a trifluoromethyl ketone (TFK) moiety and have a sulfur atom α to the ketone. In this study, sulfur-aromatic interactions were observed between the sulfur atom of OTFP and a conserved aromatic residue in the crystal structure. Mutational analysis supported the hypothesis that these interactions contribute to the potency of sulfur-containing TFK inhibitors. Together, these results clarify the binding mechanism of JHE inhibitors and provide useful observations for the development of additional enzyme inhibitors for a variety of enzymes.",
author = "Mark Wogulis and Wheelock, {Craig E.} and Kamita, {Shizuo G.} and Hinton, {Andrew C.} and Whetstone, {Paul A.} and Hammock, {Bruce D.} and Wilson, {David K.}",
year = "2006",
month = "4",
day = "4",
doi = "10.1021/bi0521644",
language = "English (US)",
volume = "45",
pages = "4045--4057",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "13",

}

TY - JOUR

T1 - Structural studies of a potent insect maturation inhibitor bound to the juvenile hormone esterase of Manduca sexta

AU - Wogulis, Mark

AU - Wheelock, Craig E.

AU - Kamita, Shizuo G.

AU - Hinton, Andrew C.

AU - Whetstone, Paul A.

AU - Hammock, Bruce D.

AU - Wilson, David K.

PY - 2006/4/4

Y1 - 2006/4/4

N2 - Juvenile hormone (JH) is an insect hormone containing an α/β-unsaturated ester consisting of a small alcohol and long, hydrophobic acid. JH degradation is required for proper insect development. One pathway of this degradation is through juvenile hormone esterase (JHE), which cleaves the JH ester bond to produce methanol and JH acid. JHE is a member of the functionally divergent α/β-hydrolase family of enzymes and is a highly efficient enzyme that cleaves JH at very low in vivo concentrations. We present here a 2.7 Å crystal structure of JHE from the tobacco hornworm Manduca sexta (MsJHE) in complex with the transition state analogue inhibitor 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP) covalently bound to the active site. This crystal structure, the first JHE structure reported, contains a long, hydrophobic binding pocket with the solvent-inaccessible catalytic triad located at the end. The structure explains many of the interactions observed between JHE and its substrates and inhibitors, such as the preference for small alcohol groups and long hydrophobic backbones. The most potent JHE inhibitors identified to date contain a trifluoromethyl ketone (TFK) moiety and have a sulfur atom α to the ketone. In this study, sulfur-aromatic interactions were observed between the sulfur atom of OTFP and a conserved aromatic residue in the crystal structure. Mutational analysis supported the hypothesis that these interactions contribute to the potency of sulfur-containing TFK inhibitors. Together, these results clarify the binding mechanism of JHE inhibitors and provide useful observations for the development of additional enzyme inhibitors for a variety of enzymes.

AB - Juvenile hormone (JH) is an insect hormone containing an α/β-unsaturated ester consisting of a small alcohol and long, hydrophobic acid. JH degradation is required for proper insect development. One pathway of this degradation is through juvenile hormone esterase (JHE), which cleaves the JH ester bond to produce methanol and JH acid. JHE is a member of the functionally divergent α/β-hydrolase family of enzymes and is a highly efficient enzyme that cleaves JH at very low in vivo concentrations. We present here a 2.7 Å crystal structure of JHE from the tobacco hornworm Manduca sexta (MsJHE) in complex with the transition state analogue inhibitor 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP) covalently bound to the active site. This crystal structure, the first JHE structure reported, contains a long, hydrophobic binding pocket with the solvent-inaccessible catalytic triad located at the end. The structure explains many of the interactions observed between JHE and its substrates and inhibitors, such as the preference for small alcohol groups and long hydrophobic backbones. The most potent JHE inhibitors identified to date contain a trifluoromethyl ketone (TFK) moiety and have a sulfur atom α to the ketone. In this study, sulfur-aromatic interactions were observed between the sulfur atom of OTFP and a conserved aromatic residue in the crystal structure. Mutational analysis supported the hypothesis that these interactions contribute to the potency of sulfur-containing TFK inhibitors. Together, these results clarify the binding mechanism of JHE inhibitors and provide useful observations for the development of additional enzyme inhibitors for a variety of enzymes.

UR - http://www.scopus.com/inward/record.url?scp=33645549367&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33645549367&partnerID=8YFLogxK

U2 - 10.1021/bi0521644

DO - 10.1021/bi0521644

M3 - Article

VL - 45

SP - 4045

EP - 4057

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 13

ER -