The Structure of Glycinamide Ribotide

The occurrence of glycinamide ribotide and its formyl derivative and the involvement of these compounds in the synthesis of inosinic acid have been discussed previously (l-4).This paper describes a method of isolating and purifying glycinamide ribotide and presents evidence that it has the structure N-glycyl-5phosphoribofuranosylamine.EXPERIMENTAL Materials-The pigeon liver extract preparation and the substrates required for the synthesis of glycinamide ribotide have been described previously (l-4).Intestinal phosphatase, purified grade, was obtained from Pentex, Inc., Kankakee, Illinois.Adenosine-5'-phosphate was obtained from the Ernst Bischoff Company, Inc., Ivoryton, Connecticut.Adenosine-3'-phosphate was obtained from the Schwarz Laboratories, Inc., New York.Methods-For paper chromatography Whatman No. 1 paper was washed with 2 N acetic acid and then with 1O-4 M ethylenediamine sodium tetraacetate.The following solvent systems were employed for paper chromatography (all ratios are in volume proportions) : (1) n-butanol-17.6N acetic acid-water(2: 1: I), (2) isopropanol-water (70:30) in an atmosphere of ammonia (0.35 ml. of concentrated ammonium hydroxide per liter of jar volume), (3) propanol-water (70: 30), (4) 1.0 M ammonium acetate, pH 7.5-95 per cent ethanol (30:75), (5) 95 per cent ethanol-O.025M potassium borate, pH 8.2 (60:40), (6) 100 gm. of phenol plus 39.5 gm. of water, (7) tertiary amyl alcohol saturated with 0.05 M potassium phthalate, pH 6.0, and (8) 1.0 M ammonium acetate, pH 7.5-95 per cent ethanol (30: 60).In