TY - JOUR
T1 - Determination of the scissile bond in the hydrolysis of α-d-ribofuranose 1-phosphate by alkaline phosphatase, acid phosphatase and formic acid and in its conversion to d-ribose 5-phosphate by phosphoglucomutase. 18O shift on the 31P-NMR and mass spectroscopic evidence
AU - Jordan, Frank
AU - Kuo, Donald J.
AU - Salamone, Salvatore J.
AU - Wang, Alice L.
N1 - Funding Information:
We are grateful to the National Institutes of Health for grant No. GM 26682, the Charles and Johanna Busch Biomedical Grant (at Rutgers) and the Rutgers Research Council for financial support. We also thank Drs. George McDonald and Dee Huang for help with the NMR spectrometer. The Middle Atlantic NMR facility at the University of Pennsylvania is supported by the National Institutes of Health Grant RR 542. We are especially grateful to Dr. William Garland of the Biochemistry and Drug Metabolism Section of Hoffmann-La Roche, Inc., Nutley, N J, for performing the mass spectral determinations.
PY - 1982/6/24
Y1 - 1982/6/24
N2 - α-d-Ribofuranose 1-[18O4]phosphate (made enzymatically employing purine nucleoside phosphorylase, inosine, and P18O4) was hydrolyzed in H216O by alkaline phosphatase, acid phosphatase, formic and hydrochloric acids. The two enzymes cleaved the OP bond, the acids the CO bond. Phosphoglucomutase formed the OP bond of D-ribose 5-phosphate when starting with α-d-ribofuranosyl 1-[18O4]phosphate. The two techniques employed in the analysis (18O shift on the 31P-NMR and mass spectrometry) were in total agreement with the above conclusions. The mass spectroscopic approach was also employed to demonstrate O-P bond cleavage by alkaline phosphatase in 5′-AMP and in d-ribose 5-phosphate. These results were obtained by hydrolysis of 16O-labelled substrate in 85 atom% H218O on a micro level (50 μl).
AB - α-d-Ribofuranose 1-[18O4]phosphate (made enzymatically employing purine nucleoside phosphorylase, inosine, and P18O4) was hydrolyzed in H216O by alkaline phosphatase, acid phosphatase, formic and hydrochloric acids. The two enzymes cleaved the OP bond, the acids the CO bond. Phosphoglucomutase formed the OP bond of D-ribose 5-phosphate when starting with α-d-ribofuranosyl 1-[18O4]phosphate. The two techniques employed in the analysis (18O shift on the 31P-NMR and mass spectrometry) were in total agreement with the above conclusions. The mass spectroscopic approach was also employed to demonstrate O-P bond cleavage by alkaline phosphatase in 5′-AMP and in d-ribose 5-phosphate. These results were obtained by hydrolysis of 16O-labelled substrate in 85 atom% H218O on a micro level (50 μl).
KW - Acid phosphatase
KW - Alkaline phosphatase
KW - Formic acid
KW - Phosphoglucomutase
KW - Ribose phosphate hydrolysis
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U2 - 10.1016/0167-4838(82)90064-4
DO - 10.1016/0167-4838(82)90064-4
M3 - Article
AN - SCOPUS:49049133213
SN - 0167-4838
VL - 704
SP - 427
EP - 436
JO - Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
JF - Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
IS - 3
ER -