Characterisation of two types of head activator receptor on hydra cells.
A head activator (HA) analogue is described which even at high concentrations does not lose its biological activity. By cross-linking two HA molecules over a C8 spacer, the conformation was sufficiently altered, such that self-inactivation of HA by dimerisation was prevented. In addition, the introduction of a tyrosine instead of phenylalanine in one of the two HA molecules allowed radioactive labelling with iodine. This HA bipeptide was used to investigate the effect of HA at different concentrations and as ligand for HA receptor characterisation. We found that low concentrations (0.1-10 pM) sufficed to stimulate interstitial cell mitosis, and that higher concentrations (10-1000 pM) were required for the determination of interstitial cells to nerve cells. Binding of the radioactive HA ligand to living hydra and to purified membrane fractions was saturable and specific. Binding was compatible with HA analogues with a stable monomeric conformation, but less well with dimerising HA and HA analogues. Scatchard and kinetic analyses revealed the presence of at least two types of binding site in the membrane fraction, one with a 'lower' affinity (Kd = 10(-9) M) and one with a 100-fold higher affinity (Kd = 10(-11) M). Autoradiography showed that interstitial cells were differentially labelled, suggesting that the number or types of HA receptors may vary depending on cell cycle status. A mutant of hydra with a multiheaded morphology contained 6-20-times more HA receptors per mg protein than other hydra species or mutants.
Neubauer
K H
KH
Center for Molecular Biology, Heidelberg, F.R.G.
Christians
S
S
Hoffmeister
S A
SA
Kreger
S
S
Schaller
H C
HC
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002453:Cell Cycle; D004151:Dipeptides / Q000138:chemical synthesis / Q000378:metabolism; D006829:Hydra / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D007700:Kinetics; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000138:chemical synthesis / Q000378:metabolism*; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D011956:Receptors, Cell Surface / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism; D018000:Receptors, Peptide; D013045:Species Specificity; D013329:Structure-Activity Relationship
D004151:Dipeptides; D009479:Neuropeptides; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; C069362:head activator receptor, Hydra; C069363:Y-head activator-head activator bipeptide; C032522:head activator peptide; D011761:Pyrrolidonecarboxylic Acid