Stabilization of vasopressin-induced membrane events by bifunctional imidoesters.
Vasopressin increases the water permeability of the luminal membrane of the toad bladder epithelial cell. This change in permeability correlates with the occurrence in luminal membranes of intramembrane particle aggregates, which may be the sites for transmembrane water flow. Withdrawal of vasopressin is ordinarily associated with a rapid reduction of water flow to baseline values and a simultaneous disappearance of the particle aggregates. The bifunctional imidoesters dithiobispropionimidate (DTBP) and dimethylsuberimidate (DMS), which cross-link amino groups in membrane proteins and lipids, slow the return of water flow to baseline after vasopressin withdrawal. Cross-linking is maximal at pH 10, and is reduced as pH is lowered. Freeze-fracture studies show persistence of luminal membrane particle aggregates in cross-linked bladders and a reduction in their frequency as water flow diminishes. Fusion of aggregate-containing cytoplasmic tubular membrane structures with the luminal membrane is also maintained by the imidoesters. Reductive cleavage of the central S-S bond of DTBP by beta-mercaptoethanol reverses cross-linking, permitting resumption of the rapid disappearance of the vasopressin effect. Bladders that have undergone DTBP cross-linking and beta-mercaptoethanol reduction respond to a second stimulation by vasopressin. Thus, the imidoesters provide a physiologic and reversible means of stabilizing normally rapid membrane events.
D000818:Animals; D001127:Arginine Vasopressin / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D002024:Bufo marinus; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007096:Imidoesters / Q000494:pharmacology*; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017483:Receptors, Vasopressin; D012964:Sodium / Q000378:metabolism; D001743:Urinary Bladder / Q000648:ultrastructure*
D003432:Cross-Linking Reagents; D007096:Imidoesters; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D017483:Receptors, Vasopressin; D001127:Arginine Vasopressin; D012964:Sodium