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Ligand Binding By A Recombinant Insect Juvenile Hormone Binding Protein, Kazushige Touhara, Keith A. Lerro, Bryony C. Bonning, Bruce D. Hammock, Glenn D. Prestwich
Ligand Binding By A Recombinant Insect Juvenile Hormone Binding Protein, Kazushige Touhara, Keith A. Lerro, Bryony C. Bonning, Bruce D. Hammock, Glenn D. Prestwich
Bryony C. Bonning
A cDNA for the hemolymph juvenile hormone binding protein (JHBP) of larval Manduca sexta has been isolated, sequenced, and expressed in an insect cell line. A recombinant baculovirus, containing the JHBP cDNA fused to the p10 promoter of Autographa californica nuclear polyhedrosis virus, was constructed. Insect cells (Sf9) infected with this virus secreted recombinant JHBP (rJHBP) into the medium (>50 pg/mL), and cotranslational removal of an 18 amino acid leader sequence was observed. rJHBP was cross-reactive with an antiserum prepared to the hemolymph JHBP and was specifically labeled by [3H]EHDA, a photoaffinity analog of JH 11, demonstrating that rJHBP …