The separation of the phosphate esters of muscle by paper chromatography

Abstract

The method of Hanes and Isherwood (1949) for separating phosphate esters by paper chromatography was used quantitatively in the study of these esters in muscle. Tests with known mixtures of phosphate esters and the recovery of the P of muscle extracts from the chromatograms suggest that the individual values obtained with the method in its present form are reliable to within 10 %. Trichloroacetic acid extracts of muscle were found to contain certain phosphate esters not yet identified. The pyrophosphate fraction present in the extracts is found to be entirely in the form of adenosine triphosphate and adenosine diphosphate. Striated muscle from tortoise, which is about 10 times slower than that from frog, contains less adenosine triphosphate and creatine phosphate plus orthophosphate than the latter. The chromatographic method was used to repeat some of the classical expts. on the metabolism of intact muscle and the results of these expts. were confirmed. An attempt was made to follow the changes in tortoise muscle during the early stages of activity. The most prominent ones are a decrease in the "high-energy" phosphate (2/3 adenosine triphosphate + 1/2 adenosine diphosphate + creatine phosphate), largely in the creatine phosphate component, and an increase in the mono-phosphate ester fraction.