A Simple Paper Chromatographic Method for Determination of Amino-Acid Activation Profiles in Tissue Preparations

Abstract

THE capacity of an enzyme preparation to activate amino-acids is usually measured by means of the amino-acid dependent isotope exchange reaction between pyrophosphate labelled with phosphorus-32 and adenosine triphosphate¹. According to the procedure usually adopted, the labelled adenosine triphosphate is separated from the other components of the incubation mixture by treatment with charcoal. The pyrophosphate of the adsorbed adenosine triphosphate is eluted by hydrolysis with hot hydrochloric acid. The radioactivity is determined and calculated per μmole adenosine triphosphate. For measurement of whole series of individual amino-acid activating enzyme activities in tissue preparations there was a need of a simple procedure which also could be used on a semi-micro scale. A scanning method was therefore developed involving paper chromatography combined with radioautography of the papergrams.