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Photoaffinity labelling of juvenile hormone binding proteins in the cockroach Leucophaeamaderae
Authors:Glenn D. Prestwich  Gae E. Kovalick  John K. Koeppe
Affiliation:Department of Chemistry State University of New York Stony Brook, NY 11794 USA;Department of Zoology University of North Carolina Chapel Hill, NC 27514 USA
Abstract:The synthesis and testing of several diazocarbonyl JH analogs (diazo JHA) which act as photoaffinity labels for insect juvenile hormone binding proteins are described. The best competitor, 10,11-epoxyfarnesyl diazoacetate, has been shown to irreversibly reduce [3H]-JH III binding to both ovarian and hemolymph JHBP from Leucophaeamaderae after irradiation at 254 nm for 20 seconds. No loss of activity was observed after incubation of JHBP and diazo JHA without irradiation. Protection from photoinactivation by diazo JHA II was achieved by the presence of an equimolar amount of JH III during the photolysis. Photoaffinity labeled proteins show loss of binding capacity without alteration of the binding affinity. This is the first example of the use of a photoaffinity label in the study of JH action on a molecular level, and may become a valuable tool in the elucidation of JH-receptor-chromatin interactions.
Keywords:DCC  dextran-coated charcoal  JH  Juvenile hormone  JHA  JH analog  JHBP  JH binding protein  TLC  thin layer chromatography  UV  ultraviolet spectroscopy  NMR  nuclear magnetic resonance spectroscopy  PEG  polyethylene glycol  PMSF  phenylmethylsulfonyl fluoride  RBA  relative binding affinity
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