Purification and characterization of the head-activator receptor from a multi-headed mutant of Chlorohydra viridissima.
In hydra the neuropeptide head activator (HA) is responsible for head-specific growth and differentiation processes. The effects of HA are mediated by high and low affinity receptors on hydra cells. In the current study HA receptors from a multi-headed mutant of Chlorohydra viridissima were solubilized from the membrane fraction using 1% Triton X-100 and 2.5 M urea. Scatchard analysis showed that the solubilized receptor had a Kd of 1.55 x 10(-9) M, indicating the low affinity subtype of the HA receptor. The solubilized receptor was purified by DMAE chromatography and subsequent affinity chromatography to homogeneity. SDS-PAGE revealed a single protein band with a molecular mass of 96 +/- 4 kDa. The native receptor eluted during gel filtration as a 113 kDa protein, and focussed with an isoelectric point of 4.8.
Center for Molecular Neurobiology, Hamburg, Germany.
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Christians
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Neubauer
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Ulrich
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006829:Hydra / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D007526:Isoelectric Point; D008970:Molecular Weight; D009154:Mutation; D011956:Receptors, Cell Surface / Q000235:genetics / Q000302:isolation & purification*; D018000:Receptors, Peptide
D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; C069362:head activator receptor, Hydra