title: Identification of specific interaction of juvenile hormone binding protein with isocitrate dehydrogenase.
abstract: Juvenile hormone (JH) is essential for multiple physiological processes: it controls larval development, metamorphosis and adult reproduction. In insect hemolymph more than 99 % of JH is bound to juvenile hormone binding protein (JHBP), which protects JH from degradation by nonspecific hydrolases and serves as a carrier to supply the hormone to the target tissues. In Galleria mellonella hemolymph, JHBP is found in a complex with lipid-binding high molecular weight proteins (HMWP) and this interaction is enhanced in the presence of JH. In this report, we present studies on the interaction of JHBP with low molecular weight proteins (LMWP) in the hemolymph. Using ligand blotting we found that JHBP interacts with a protein of about 44 kDa. To identify the protein that preferentially binds JHBP, a LMWP fraction was applied to a Sepharose-bound JHBP and, after washing, the column was eluted with free JHBP acting as a specific competitor or with carbonic anhydrase as a negative control. The eluted proteins were separated by SDS/PAGE and analyzed by mass spectrometry. Isocitrate dehydrogenase was identified as a component of the supramolecular complex of JHBP with hemolymph proteins.
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, Wroclaw University of Technology, Wrocław, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Zalewska
authors.forename: Andrzej
authors.initials: A
authors.lastname: Ożyhar
authors.forename: Marian
authors.initials: M
authors.lastname: Kochman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006458:Hemolymph / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D007521:Isocitrate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D007605:Juvenile Hormones / Q000378:metabolism*; D007915:Lepidoptera / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid
chemical_list: D002352:Carrier Proteins; D019476:Insect Proteins; D007605:Juvenile Hormones; D007521:Isocitrate Dehydrogenase