The Path of Carbon in Photosynthesis. XVIII The Identification ofNucleotide Coenzymes Page: 4 of 24
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UCRL-2074
authentic adenosine, and by enzymatic deamination to inosine, identified by
co-chromatography and by its hydrolysis to give hypoxanthine and ribose. The
phosphatase preparation used for enzymatic dephosphorylation was found to con-
tain a deaminase.
Experimental Part
When the plant extract is chromatographed as a band in a unidimensional
chromatogram (phenol solvent) and the slow-moving u.v.-absorbing band extracted
and hydrolyzed. with acid, at least three uov.-absorbing compounds are produced.
These are adenine, adenosine-5 -phosphate and uridine-5'-phosphate. All three
compounds have been characterized by their u.v. absorption spectra and Rf values
in various solvents. The location of the phosphate group in the uridylic acid
has been shown by its reaction with sodium periodate. The adenylic acid has
the same Rf value as adenosine-5'-phosphate (muscle adenylic acid) and dif-
ferent from that of yeast adenylic acid.
The radioactive compounds produced when the original is subjected to
rechromatography have been examined. In particular, the two spots which yielded
only glucose and galactose on dephosphorylation were subjected to further study.
They had properties very reminiscent of the "fast ester" and "slow ester" of
Leloir and his co-workers,3 which have been shown to be, respectively, glucose-
1,2-cyclic phosphate and a mixture of glucose-l- and glucose-2-phosphates. The
corresponding phosphates of galactose have very similar properties. Glucose-
1,2-cyclic phosphate was prepared by the action of ammonia on uridine diphosphate
glucose, and co-chromatographed with the suspected "cyclic phosphate" area, ie.
that with 'I values nearly as great as those of glucose itself.
We have attempted to chromatograph the original unknown in the ethanol/
ammonia solvent of Paladini and Leloir.3 It has not been possible to d emon-
strate quantitative conversion into cyclic phosphates, probably due to the-4-
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Buchanan, J. G.; Lynch, V. H.; Benson, A. A.; Calvin, M. & Bradley, D. F. The Path of Carbon in Photosynthesis. XVIII The Identification ofNucleotide Coenzymes, report, January 19, 1953; Berkeley, California. (https://digital.library.unt.edu/ark:/67531/metadc879339/m1/4/: accessed April 17, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.