Anti-phosphoglucomutase (PGM) antibodies have been produced by immunising a sheep with a purified preparation of rabbit skeletal muscle PGM and used to devise an immunological procedure for detecting PGM isozymes after isoelectric focusing. The anti-rabbit PGM antibodies cross react with human PGM and can be used to identify the PGM1 isozymes characteristic of this polymorphism. The patterns revealed by immunodetection are exactly comparable with those obtained by isozyme staining.

Animals↗

PubMedDate not supplied

Biochemical genetic analysis of human and rodent aldehyde dehydrogenase (ALDH).

ALDH isozymes have been characterized in terms of substrate and coenzyme specificity, heat stability, tissue distribution and electrophoretic properties. The activity of the isozymes has also been examined in rodent-human somatic cell hybrids in order to map the structural genes to specific chromosomes and to study the control of gene expression. One isozyme, designated ALDH3, which is very active against benzaldehyde, was found to show variable expression in hybrids made between rat hepatoma cells and human fibroblasts or fetal liver. Segregation analysis of these hybrids indicates that the structural locus for human ALDH3 may be on chromosome 17. The expression of rodent ALDH3 in these hybrids was extremely variable and not correlated with the appearance of the human enzyme. In hybrids expressing human and rodent ALDH3 no heteromeric isozymes were observed. The human "cytosolic" ALDH1 and "mitochondrial" ALDH2 isozymes did not appear to be expressed in any of the somatic cell hybrids examined.

Aldehyde Dehydrogenase↗

PubMedDate not supplied

Immunological reactivity of native and modified alcohol dehydrogenase (ADH).

The extent to which treatment of horse liver alcohol dehydrogenase (ADH) by procedures known to disturb the Zn association induced conformation changes detectable by immunological techniques has been investigated. Treatment of ADH by sodium dodecyl sulphate or by total reduction and carboxymethylation leads to complete loss of reactivity with a rabbit immune serum against the native enzyme. After selective carboxymethylation the enzymatic activity was reduced but the preparation had the same immunological activity of the native enzyme. Similar results were obtained when ADH was treated with reagents known to react with the "functional" Zn atoms, such as sodium dietyldithiocarbamate, 1,10 phenanthroline, and 2,2' bipyridine. In contrast dialysis in 0.01M phosphate buffer containing 0.1mM EDTA removing the "structural" Zn atoms leads to a parallel decrease of enzymatic and immunological activity. Thus loss of "structural" Zn atoms affects the immune reactivity of ADH differently from the loss of "functional" Zn atoms.

Alcohol Dehydrogenase↗