Retention of a functional resact receptor in isolated sperm plasma membranes.
Resact, a peptide obtained from eggs, causes a change in the Mr, and a loss of 32P from a plasma membrane protein identified as guanylate cyclase. Here, a resact analog (125I-[Tyr1, Ser8] resact) was synthesized and shown to bind to isolated sperm membranes. Resact, but not speract, competed with the radiolabeled ligand for binding. When membranes were prepared under appropriate conditions, guanylate cyclase remained at Mr 160,000; the incubation of membranes with gamma-32P-ATP resulted in the formation of 32P-labeled guanylate cyclase. The addition of resact to the membranes caused a shift in the Mr, a complete loss of 32P, and a 70% reduction in guanylate cyclase activity within 1 min; resact had an ED 50 at 100 nM concentration. Speract failed to cause any of these effects. This represents the first demonstration of receptor-mediated responses of isolated sperm membranes identical to those seen in the intact cell.
Bentley
J K
JK
Shimomura
H
H
Garbers
D L
DL
D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism*; D002633:Chemotaxis; D006152:Cyclic GMP / Q000378:metabolism; D004527:Egg Proteins; D006162:Guanylate Cyclase / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D010766:Phosphorylation; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012617:Sea Urchins / Q000378:metabolism; D013094:Spermatozoa / Q000032:analysis*
D004527:Egg Proteins; D008565:Membrane Proteins; D009842:Oligopeptides; D010455:Peptides; D011956:Receptors, Cell Surface; C028364:speract; C042019:resact; D006162:Guanylate Cyclase; C048639:resact protein, sea urchin; D006152:Cyclic GMP