J. Biochem, 1976, Vol. 80, No. 1 27-32
© 1976 Japanese Biochemical Society
research-article |
Synthesis of Various Phosphodiesters and Phosphomonoesters with Ribonuclease N1
Department of Biophysics and Biochemistry, Faculty of Science, The University of Tokyo Hongo, Bunkyo-ku, Tokyo 113 Department of Agricultural Chemistry, Faculty of Agriculture, The University of Tokyo Hongo, Bunkyo-ku, Tokyo 113
1. 3'-Guanylyl-ethanol, 3'-guanylyl-propanol, and 3'-guanylyl-
-glycerol were synthesized by ribonuclease N1 [EC 3. 1. 4. 8] using guanosine 2', 3'-cyclic phosphate as a phosphate donor and various alcohols as phosphate acceptors. The yields of these phosphodiesters were 15%, 13.5%, 38.2%, respectively, with respect to phosphate donor under the optimum conditions. No phosphodiester was synthesized when 2-propanol was used as a phosphate acceptor. Thus, primary alcoholic hydroxyl groups may be regarded as the preferred phosphate acceptor.
2. 3'-Guanylyl-glucose and 3'-guanylyl-ribose were synthesized using glucose and ribose as phosphate acceptors. Under the optimum conditions, the yields of guanylyl-glucose amounted to 52.0%, while that of guanylyl-ribose was much lower. The guanylyl-glucose can be regarded as 3'-guanylyl-6-glucopyranose, based on the results of periodate oxidation.
3. Neither hydroxyamino acids (serine and threonine) nor N-acetylserinamide could be phosphorylated under the conditions used for the above phosphorylations.
4. 3'-Guanylyl-glycerol obtained as above was hydrolyzed by snake venom phospho-diesterase to produce glycerol 3-phosphate. The latter consisted of L-glycerol 3-phosphate (ca. 17%) and the D-isomer (ca. 83%). Ribonuclease N1 thus catalyzes an asymmetric synthesis.
1Present address: The Institute of Molecular Biology, Faculty of Science, Nagoya University Nagoya, Aichi 464.
2Present address: Mitsubishi-Kasei Institute of Life Sciences Minamiooya, Machida-shi, Tokyo 194.