Effect of the esterase-D phenotype on its in vitro enzyme activity

Esterase-D phenotypes and in vitro activity have been measured in red blood cells from 258 retinoblastoma patients and 73 unaffected relatives. Individuals with the 1-1 and 2-1 phenotypes showed distributions of enzyme activity which were not significantly different from each other. Individuals with the 2-2 phenotype, however, consistently showed a 25-30% lower level of enzyme activity. These results demonstrate the importance of determining the esterase-D phenotype in individuals with low ESD activity who might otherwise be assumed to carry a chromosome deletion at the esterase-D locus. We have also shown that, in vitro, the ESD enzyme is unstable over relatively short periods of time which, if uncontrolled, can give rise to a large variation in measured enzyme levels. The addition of b-mercaptoethanol to the assay buffer, which stabilises the enzyme, results in more consistent values being obtained within the same ESD phenotype. This feature could account in part for much of the variability in enzyme activity observed between different individuals in other studies.