Characterization of affinity-purified juvenile hormone esterase from Trichoplusia ni.

T. N. Hanzlik, B. D. Hammock

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Abstract

Juvenile hormone (JH) esterase was purified greater than 1000-fold in one step from hemolymph and whole larval homogenates from the last larval instar of Trichoplusia ni to give a single diffuse band that migrates at Mr = 64,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purification was based on an affinity chromatography procedure that employs trifluoromethyl ketone ligands. Isoelectric focusing of the purified preparations resulted in multiple bands that coincided to all significant hydrolysis of juvenile hormone detected in this manner. Kinetic experiments using optically pure enantiomers of JH II as substrates showed the two main electromorphs of JH esterase from the hemolymph to have apparently identical kinetic parameters as well as a similar capability to distinguish between substrates that differ in the orientation of the epoxide moiety of JH. However, the enzyme could hydrolyze esters lacking the JH structure. The proteins were shown to be monomers and to have asparagine-linked oligosaccharides, most likely of hybrid structure. Immunochemical and other evidence showed that the affinity-purified proteins were responsible for all significant JH esterase activity during periods of rapid esterolysis in vivo.

Original languageEnglish (US)
Pages (from-to)13584-13591
Number of pages8
JournalJournal of Biological Chemistry
Volume262
Issue number28
StatePublished - Oct 5 1987

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Juvenile Hormones
Hemolymph
Affinity chromatography
Enantiomers
Asparagine
Epoxy Compounds
Isoelectric Focusing
Substrates
Ketones
Electrophoresis
Oligosaccharides
Affinity Chromatography
Kinetic parameters
Sodium Dodecyl Sulfate
Purification
Polyacrylamide Gel Electrophoresis
Hydrolysis
Esters
Proteins
Monomers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of affinity-purified juvenile hormone esterase from Trichoplusia ni. / Hanzlik, T. N.; Hammock, B. D.

In: Journal of Biological Chemistry, Vol. 262, No. 28, 05.10.1987, p. 13584-13591.

Research output: Contribution to journalArticle

Hanzlik, T. N. ; Hammock, B. D. / Characterization of affinity-purified juvenile hormone esterase from Trichoplusia ni. In: Journal of Biological Chemistry. 1987 ; Vol. 262, No. 28. pp. 13584-13591.
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