Identification, Development, and Regional Distribution of Thymidylate Synthetase in Adult Rabbit Brain

Abstract

Development and regional distribution of thymidylate synthetase (TS) (EC 2.1.1.45) in rabbit brain were determined. After optimization of the assay for brain, TS activity in brain was measured by a nonspecific (3H2O release) and specific method. The specific method involved the conversion of [6-3H]deoxyuridine monophosphate (dUMP) to [3H]thymidine phosphate and the subsequent identification of [3H]thymidine. The specific activity of the enzyme in whole brain of newborn rabbits declined from 10.35 .+-. 1.17 units/mg protein to 0.71 .+-. 0.09 units/mg protein at 10-12 wk of age. Two yr old rabbits had 0.81 .+-. 0.04 units/mg protein. The decline in specific activity with age was not due to an inhibitor of TS activity or a change in the Km for dUMP. The Km for dUMP of the unpurified enzyme in the brains of both 10 day old and young adult rabbits was 0.8 .mu.M. In young adult rabbits (3 mo.) the specific activity of TS was similar in the various regions of the brain tested except for the cerebellum, which had 40% higher specific activity than the whole brain. TS is widely distributed in adult rabbit brain, and, although the activity declines with age, it stabilizes at adult levels at 3 mo.