Specific binding sites for 125I-labelled human luteinizing hormone (hLH) in microsomal fractions from Drosophila.
Microsomal fractions prepared from Drosophila flies and larvae bound 125I-labelled human luteinizing hormone (hLH) in a displaceable manner. Binding increased linearly with increasing microsome concentration, and was dependent on the pH and metal ion concentration of the medium, and on the temperature and duration of incubation. Scatchard plots of 125I-hLH binding demonstrated the presence of two distinct binding sites, one with high affinity and low capacity, the other with low affinity and high capacity. 125I-hLH binding was inhibited in a dose-dependent fashion by partially purified human gonadotropin preparations, but not by other proteins, hormones and peptides. Highly purified human chorionic gonadotropin (hCG) or hLH preparations were much less effective at inhibiting 125I-hLH binding to Drosophila microsomes than partially purified gonadotropins. This was due to the presence of a contaminant present in partially purified commercial hCG preparations. Chromatography of crude hCG preparations clearly resolved the LH-binding inhibitor from hCG. Moreover, whereas hCG (and epidermal growth factor-like factors present in crude hCG preparations) were not bound to CM-Sepharose, the Drosophila LH-binding inhibitor was adsorbed strongly. Fractionation of 125I-hLH tracer on concanavalin A-Sepharose suggested that Drosophila binding sites did not preferentially bind a subfraction of the hormone tracer. These data suggest that high affinity binding sites for hLH/hCG-like molecules are present in Drosophila microsomes.
Bramley
T A
TA
University of Edinburgh Department of Obstetrics and Gynaecology, Scotland, U.K.
Menzies
G S
GS
Jowett
T
T
D000818:Animals; D001667:Binding, Competitive; D002413:Cations, Divalent / Q000494:pharmacology; D006063:Chorionic Gonadotropin / Q000378:metabolism; D004331:Drosophila melanogaster / Q000254:growth & development / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007814:Larva; D007986:Luteinizing Hormone / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D011974:Receptors, LH / Q000302:isolation & purification / Q000378:metabolism*; D013045:Species Specificity; D013696:Temperature
D002413:Cations, Divalent; D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D007986:Luteinizing Hormone