Judith J. Britten

Studies on the inhibition by propranolol of some human erythrocyte membrane enzymes and plasma cholinesterase

Human erythrocyte acetylcholinesterase and the plasma cholinesterase variants are not only inhibited by propranolol but have been found to show stereospecificity for its isomers. The erythrocyte enzyme has a greater affinity for the L-isomer than either the racemate or the D-isomer. In contrast the plasma cholinesterases have greater specificity for the D-isomer than the other isomer or racemate. The usual enzyme shows greater stereospecificity than the atypical enzyme and these findings present additional evidence that these enzyme variants differ in structure at the catalytic active site. Neither Na+ + K+ -ATPase nor Mg2+-ATPase show stereo-specificity for the isomers of propranolol although both enzymes are inhibited by the drug. The action of the drug on the four enzymes in blood samples obtained from patients having Huntingtons disease was found to be identical to those observed on the enzymes in blood samples from healthy controls.

Inhibition of the plasma cholinesterase variants by pancuronium bromide and some of its analogues

Pancuronium bromide, a 3,17-diacetoxy-5 alpha-androstane, and three of its analogues, the 17-desoxy, the 3,17-dibutyryloxy and the 16-N-monoquaternary ammonium derivatives have been used as inhibitors of the usual and atypical plasma cholinesterase variants. In all cases the usual enzyme is more sensitive to inhibition by the substituted steroids than the dibucaine resistant enzyme. The relative affinities of the bis-quaternary ammonium compounds for either enzyme is in the order dibutyryloxy derivative > 17-desoxy derivative greater than or equal to pancuronium bromide. The monoquaternary compound has the least affinity of all the inhibitors for the usual enzyme but the greater affinity for the atypical enzyme. These observations show that the bis-quaternary compounds are very powerful differentiators of the variants. The monoquaternary derivative shows less differential inhibition, but provides additional evidence that the usual and dibucaine resistant variants differ in structure at or near their esteratic active site.

Recognition of Two New Phenotypes Segregating the E1k Allele for Plasma Cholinesterase

The first identification of the cholinesterase variants E1kE1k and E1kE1s is reported from a family study. The evidence is based on the biochemical parameters of enzymic activity, and dibucaine, fluoride and RO2 numbers. Two individuals appear to be homozygotes E1kE1k and two are heterozygotes E1kE1s with family evidence supportive of these genotypes. The heterozygotes E1kE1s will be sensitive to suxamethonium.

Differential Inhibition of Plasma Cholinesterase Variants Using the Dibutyrate Analogue of Pancuronium Bromide

A steroid, the dibutyrate analogue of pancuronium bromide (9.8 X 10(-8)M), has been used as differential inhibitor in the study of the plasma cholinesterase variants. Pancuronium dibutyrate numbers have been measured for 190 individuals, and the mean values for six of the known genotypes, E1uE1u, E1uE1f, E1uE1a, E1fE1a, E1aE1a, and E1fE1f, have been calculated. Evidence is presented that a combination of the pancuronium dibutyrate number and the fluoride number give better resolution of the six genotypes than the combination of the pancuronium dibutyrate and the dibucaine number. This new differential inhibitor has real potential for revealing the probable existence of new genotypes.

A comparison of some methods of phenotyping the plasma cholinesterase variants using benzoylcholine as substrate.

Five differential inhibitors of plasma cholinesterase have been compared using benzoylcholine as substrate. None of the inhibitors (dibucaine, NaF, NaBr, NaCl, or pancuronium dibutyryloxy bromide) could be used singly to resolve all the variants. Better resolution was obtained when two inhibitors were used in conjunction. Clear differentiation of all six genotypes was obtained only with the combined use of pancuronium dibutyryloxy bromide and sodium fluoride. The limitations of some of the parameters are discussed.

Segregation of the E1J gene for plasma cholinesterase in family studies

Four families are reported in which the E1j gene is segregating. Two E1jE1k and one E1jE1f genotypes have been recognised by genetic analysis. The biochemical characteristics of several E1j genotypes are presented.

Recognition of the Ek1Ek1 homozygote for plasma cholinesterase.

A family is reported in which the propositus was found to be sensitive to suxamethonium. Both parents were heterozygote each having genotype Ea1Ek1. The sibling of the propositus had the most common phenotype as defined by dibucaine, fluoride and RO2 numbers. Genetic analysis, however, indicated that this sibling must be an Ek1Ek1 homozygote.