Abstract
AbstractJuvenile hormone epoxide hydrolase (JHEH) and juvenile hormone diol phosphotransferase (JHDPT) were characterized from the Malpighian tubules of day 1 fifth instar Manduca sexta. An improved RP‐HPLC assay is described for the major metabolites of (10R, 11S) juvenile hormone I: diol, acid, aciddiol, and diol‐phosphate. JHEH is strictly associated with membrane fractions, while JHDPT is cytosolic. JHEH may be solubilized in active form by the nonionic detergents Thesit or MEGA‐8. Separation of Malpighian tubule cytosol proteins using preparative isoelectric focusing yields two zones which contain JHDPT activity, at pl 4.8–5.1 and 6.8–8.2. The partially purified JHDPT from either zone requires both ATP and Mg2+ for activity, so this enzyme may be formally called either ATP:juvenile hormone diol phosphotransferase or juvenile hormone diol kinase (EC 2.1.7.3.). Metabolites more polar than JH I aciddiol and JH I diol‐phosphate are generated in vivo from either [3H]JH I or [3H]JH I diol. © 1995 Wiley‐Liss, Inc.
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