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Purification and characterization of the head‐activator receptor from a multi‐headed mutant of chlorohydra viridissima
Author(s) -
Christians Stefan,
Neubauer Karl-Heinz,
Ulrich Henning
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81203-c
Subject(s) - mutant , activator (genetics) , receptor , biology , microbiology and biotechnology , genetics , gene
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 K d of 1.55 × 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.