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Assay methods for methyl farnesoate esterases in crustaceans
Author(s) -
Homola Ellen,
Chang Ernest S.
Publication year - 1997
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/(sici)1520-6327(1997)36:2<115::aid-arch4>3.0.co;2-u
Subject(s) - esterase , biology , biochemistry , hepatopancreas , juvenile hormone , substrate (aquarium) , chromatography , insect , metabolism , midgut , crustacean , enzyme , chemistry , larva , hormone , botany , ecology
Methyl farnesoate (MF) is a sesquiterpenoid that is synthesized by crustaceans and is structurally similar to the insect juvenile hormones. MF is metabolized to farnesoic acid by carboxylesterases present in crustacean tissues. In order to investigate the biological significance of MF metabolism, we have developed two rapid methods for measuring MF esterase activity. The first method is a radiochemical partition assay that utilizes an authentic substrate. The [ 3 H]MF partition assay was used to evaluate the spectrophotometric esterase substrates methyl 1‐heptylthioacetothioate (HEPTAT) and methyl 1‐pentylthioacetothioate (PENTAT) for use with crude and partially purified samples of MF esterases from lobster hepatopancreas (midgut gland). The spectrophotometric method is less specific for MF esterases than the partition assay but is less time consuming and nonradioactive, and it provides kinetic information. HEPTAT and PENTAT were suitable for rapid screening of chromatographic fractions for MF esterase activity. Arch. Insect Biochem. Physiol. 36:115–128, 1997. © 1997 Wiley‐Liss, Inc.