COMPARATIVE BIOCHEMICAL STUDIES ON NORMAL AND ON POLIOMYELITIS INFECTED TISSUE CULTURES: III. ENZYME ASSAYS ON HOMOGENATES OF SURVIVING NORMAL RHESUS KIDNEY. EFFECT OF SYNTHETIC NUTRIENT MIXTURES

Abstract

Investigation of enzyme systems in normal tissues is the main subject of this communication. The activity of phosphatases, nucleotidases, and nucleases was explored in "brei" prepared with water, physiological salt mixtures, and synthetic media used for tissue cultivation, to compare the influence of these diluents. The depression of acid phosphatase by medium 597, noted previously in cells growing in vitro, was confirmed in fresh tissue and definitely proved on purified enzyme. Alkaline phosphatase was activated by the same synthetic medium. Differences in kinetics were observed when the effect of medium 597 was examined on 5-nucleotidase concentrate of Russel's viper venom and on pentanucleotidase of fresh kidney. Crystalline RNA-se was inhibited, in a manner similar to the nucleases of kidney extracts. Both phosphomonoesterases were enhanced by the less complex nutrient 697. The deterioration of DNA-se in medium 199 upon storage was striking. Significant observations were gathered on enzymes of concentrated and very diluted (1: 1600) tissue homogenates, on extracts, and on isolated enzymes of various grades of purity.