Purification and characterization of juvenile hormone esterase from hemolymph of the Colorado potato beetle

Archives of Insect Biochemistry and Physiology
A M VermuntC A de Kort

Abstract

In the Colorado potato beetle (Leptinotarsa decemlineata), low juvenile hormone (JH) titers are necessary to initiate metamorphosis and diapause. Low JH titers coincide with high activities of JH esterase, which occur mainly in the hemolymph. The specific activity of JH esterase appeared to be highest in the last larval instar, at day 3 after the molt, and reached a value of 13.5 nmol/min/mg. JH esterase was purified from hemolymph collected at this stage be a sequence of separation systems, including preparative nondenaturing PAGE, isoelectric focusing, and SDS-PAGE. The enzyme had a molecular weight of 120,000 and was composed of two subunits with molecular weights of 57,000, which were not linked by disulphide bridges. Isoelectric focusing revealed two forms of the enzyme with isoelectric points of 5.5 and 5.6. The Km and kcat of the purified enzyme were determined. The major form with pl 5.6 had a Km of 1.4 x 10(-6) M and a kcat of 0.9 s-1 and the minor form with pl 5.5 had a Km of 2.2 x 10(-6) M and a kcat of 1.9 s-1. The quaternary structure of L. decemlineata JH esterases, as differs from JH esterases in other species, which are monomers.

References

Oct 1, 1977·Analytical Biochemistry·B D Hammock, T C Sparks
Jul 15, 1976·Life Sciences·S J Kramer, C A de Kort
Dec 1, 1992·The International Journal of Biochemistry·V K WardB D Hammock
Jan 1, 1992·Archives of Insect Biochemistry and Physiology·B KoopmanschapS de Kort
Jan 1, 1994·Developmental Genetics·V VenkataramanG Jones

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Citations

Nov 6, 2003·Insect Biochemistry and Molecular Biology·Shizuo G KamitaBruce D Hammock
Mar 21, 1998·Insect Biochemistry and Molecular Biology·A M VermuntC A de Kort
Aug 27, 1998·Insect Biochemistry and Molecular Biology·P M CampbellM J Healy
Jan 5, 2000·Insect Molecular Biology·J P HajósJ M Vlak
Nov 21, 2007·Journal of Insect Physiology·H Frederik Nijhout, Michael C Reed
Jul 14, 2007·Insect Biochemistry and Molecular Biology·Hua BaiSubba Reddy Palli
May 21, 2005·Toxicon : Official Journal of the International Society on Toxinology·Richard D CroslandHarry B Hines
Jan 9, 2007·Toxicon : Official Journal of the International Society on Toxinology·Richard H ffrench-ConstantNicholas R Waterfield
Dec 26, 2001·Archives of Insect Biochemistry and Physiology·Anthony J ZeraLawrence Harshman
Sep 2, 1998·Insect Molecular Biology·A M VermuntC A de Kort
Jun 14, 2000·Insect Biochemistry and Molecular Biology·B A ThomasB D Hammock
May 14, 2005·Journal of Insect Physiology·Damu R KethidiSubba R Palli

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