By V. Z. Gorkin
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Extra info for Amine Oxidases in Clinical Research
For the assay of monoamine oxidase activity in human platelets, tyramine, tryptamine and benzylamine are the usual substrates supplied. In some studies dopamine, serotonin, 3~phenylethylamine, octopamine, 3-iodobenzylamine were also used (Murphy, 1978a). In studies using platelets direct radio metric methods of assay are used routinely. The radioactivity of the deamination products (mainly aldehydes) is measured after separating an excess of the amine by means of columns containing the anionite Amberlite CG-50 (Tipton and Youdim, 1976).
By means of antibodies to pig plasma benzylamine oxidase labelled with fluorescent dyes, proteins possessing identical antigenic determinants were found in connective tissue of aorta, uterus, kidney and tendons (Buffoni and Deila Corte, 1972). Using other techniques enzymes resembling the soluble amine oxidases of serum were also detected in connective tissue of skin, bone and cartilage, aorta and dental pulp (Nakano and coworkers, 1974). Connec tive tissue fibroblasts are considered to be a possible source of plasma amine oxidases (Nagatsu and coworkers, 1972).
A suspension of submitochondrial particles (5 mg protein/ml) were centrifuged (120 min at 144,000 g) within 30 min after adding Cutscum ( 2 % ) . Extraction of the mono amine oxidases into the supernatant was not characterized quantitatively but rapid inactivation of the enzyme was observed in the presence of Cutscum. Rat and rabbit liver mitochondrial monoamine oxidases (substrates kynuramine, tyramine and serotonin) have been solubilized by combined treatment with an anionic surface-active agent, sodium dodecylsulfate (SDS), and Cutscum.