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

Frank Jordan, Donald J. Kuo, Salvatore J. Salamone, Alice L. Wang

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Abstract

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).

Original languageEnglish (US)
Pages (from-to)427-436
Number of pages10
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume704
Issue number3
DOIs
StatePublished - Jun 24 1982

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

Keywords

  • Acid phosphatase
  • Alkaline phosphatase
  • Formic acid
  • Phosphoglucomutase
  • Ribose phosphate hydrolysis

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