n-Alkanols and halothane inhibit red cell anion transport and increase band 3 conformational change rate.
The effects of halothane and n-alkanols on band 3, the anion-exchange protein of the red cell membrane, have been characterized by radioactive sulfate exchange and equilibrium and kinetic binding of a fluorescent anion transport inhibitor, 4,4'-dibenzamido-2,2'-stilbenedisulfonic acid (DBDS), with fluorescence and stopped-flow techniques. Ethanol, butanol, hexanol, heptanol, octanol, and decanol inhibit radioactive sulfate efflux from red blood cells in a dose-dependent manner with an average Hill coefficient of 1.3 +/- 0.1. Over a 10(4)-fold range of buffer concentrations, the calculated membrane alkanol concentrations at which anion transport rates are reduced by 50% are 100-200 mM. At 100-300 mM membrane concentrations, halothane and the n-alkanols increase the apparent rate of DBDS binding to band 3 2-3-fold. Analysis of kinetic and equilibrium DBDS binding data shows that these drugs increase the rate of the DBDS-induced conformational change in the DBDS-band 3 complex. Equilibrium DBDS binding studies reveal differences between the actions of short-chain alkanols (ethanol and butanol) and those of long-chain alkanols (hexanol and longer). Short-chain alkanols reduce the equilibrium affinity of DBDS for band 3, while long-chain alkanols have no effect on equilibrium DBDS binding. The results for halothane and long-chain alkanols suggest a nonspecific, lipid-mediated mechanism of anesthetic action, which may be coupled to protein inactivation by an increase in the rate of protein conformational changes resulting in nonfunctional states. The results for short-chain alkanols indicate that they have the same nonspecific actions as the long-chain alkanols but also have specific effects on the stilbene binding site of band 3.
D000438:Alcohols / Q000494:pharmacology*; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D027321:Anion Transport Proteins; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000097:blood*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D006221:Halothane / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D013329:Structure-Activity Relationship
D000438:Alcohols; D001457:Anion Exchange Protein 1, Erythrocyte; D027321:Anion Transport Proteins; D002352:Carrier Proteins; D006221:Halothane