Characterization of hemolymph juvenile hormone esterase from Lymantria dispar |
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| Institution: | 1. School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, PR China;2. Ministry of Education and Shanghai Key Laboratory of Children’s Environmental Health Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, PR China;3. Epidemiology Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA;4. UNDP/UNFPA/UNICEF/WHO/World Bank Special Programme of Research, Development and Research Training in Human Reproduction (HRP); Department of Reproductive Health and Research, World Health Organization, Geneva 1202, Switzerland;1. Department I of Thoracic Surgery, Hospital of Nanjing Medical University, Jiangsu, China;2. Department of Thoracic Surgery, Affiliated Jiangning Hospital of Nanjing Medical University, Jiangsu, China;1. Department of Biochemistry, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Bangkok 10900, Thailand;2. Aquatic Molecular Genetics and Biotechnology Laboratory, Agricultural Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani 12120, Thailand;3. Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand |
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| Abstract: | A major peak of juvenile hormone esterase (JHE) activity approaching 330 nmol JH III hydrolyzed/min/ml of hemolymph was observed during the last larval growth stage in Lymantria dispar. A smaller peak of JHE occurred 3–5 days after pupation. The gypsy moth JHE was purified from larval hemolymph using a classical approach. A specific activity of 766 units per mg of protein and a Km of 3.6 × 10?7 M for racemic JH III and the (10R, 11S) enantiomer of JH II was determined for the purified enzyme. The 62 kDa esterase was insensitive to inhibition by O,O-diisopropyl phosphorofluoridate (DFP), or by phenylmethylsulfonyl fluoride (PMSF). Two forms of JHE isolated by RP-HPLC were indistinguishable by HPLC tryptic peptide mapping and share an identical N-terminal amino acid sequence. Polyclonal antisera raised against gypsy moth enzyme cross-reacted with JHE from Trichoplusia ni but not with JHE from Manduca sexta. A weak cross-reactivity was observed with JHE from Heliothis virescens. Forty amino acid residues of the N-terminus were placed in sequence. The N-terminal sequence of JHE from L. dispar showed little homology to the sequence of JHE from H. virescens. The immunological and structural data support the conclusion that markedly different esterases, which catalyze the hydrolysis of juvenile hormone, are present in the hemolymph of different Lepidoptera. |
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