I. Phosphorus-Containing Compounds in Neurospora. II. Synthetic Substrates for Chymotrypsin

Author: Ellman, George Leon

Year: 1952

Degree: Dissertation (Ph.D.)

Advisor: Mitchell, Herschel K.

Committee Member: Unknown, Unknown

Option: Chemistry; Biology

DOI: 10.7907/FZ0J-HB93

Abstract

I. Phosphorus-containing Compounds in Neurospora

A paper chromatographic method is described for the separation of the water soluble phosphorus compounds found in tissue extracts. Results with known compounds are given. The method has been applied to the separation of the phosphorus compounds in extracts of Neurospora. The presence of the following compounds is indicated: the adenosine phosphates, 3-phosphoglycerate, glucose-1-phosphate, glucose-6-phosphate, and fructose-6-phosphate. The presence of at least one major unknown compound is also indicated. The significance of these results is discussed.

Modifications of the usual methods for the study of the phospholipids were also devised. The phospholipids were separated into the lecithins and cephalins. The cephalins were converted into the N-2,4-dinitrophenyl derivatives. These were chromatographed on silicic acid-celite columns. The fractions collected were hydrolyzed. Titration of the fatty acids indicate the presence of some phospholipids which do not correspond to the classical formulae. Chromatography of the phosphate esters present in these hydrolyzates revealed glycerophosphate and one unknown phosphorus compound. Chromatography of the DNP-amines showed the presence of eight components. The DNP derivatives of ammonia, ethanolamine, serine, and 1-amino-2-methyl-2-propanol were identified. The techniques used and the significance of the results are discussed.

II. Synthetic Substrates for Chymotrypsin

A general synthesis of methyl acyl-phenylalanyl-anthranilates has been devised. The acetyl derivative serves as a substrate for chymotrypsin. Methyl anthranilate is produced when this compound is hydrolyzed and it is detected by its fluorescence. The succinyl and maleyl derivatives were also prepared, but they are not hydrolyzed by the enzyme. These results are discussed.

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