Abstract
The diastereomers of uridyl-(3'-5')adenyl-O,O-phosphorothioate [Up(S)A] have been separated by high-performance liquid chromatography. Their identification as RP and SP follows from the RNase A digestion of these products. It was then shown, by the same method, that the R isomer is hydrolyzed by snake venom phosphodiesterase (PDEase) approximately 500 times faster than the S isomer. Similarly, the stereoisomer of adenosine 5'-O-(1-thiotriphosphate) (ATPalphaS), until now arbitrarily designated as isomer B, is hydrolyzed ca 400 times faster by PDEase than is isomer A. From these results it is concluded that the R isomers of Up(S)A and ATPalphaS, isomers B, have the same absolute configuration. It then follows that isomer A of ATPalphaS, the preferred of the two isomers as substrate for DNA-dependent RNA polymerase, has the S configuration. The implications for the stereochemistry of action of the latter enzyme are discussed.
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Selected References
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- Eckstein F., Armstrong V. W., Sternbach H. Stereochemistry of polymerization by DNA-dependent RNA-polymerase from Escherichia coli: an investigation with a diastereomeric ATP-analogue. Proc Natl Acad Sci U S A. 1976 Sep;73(9):2987–2990. doi: 10.1073/pnas.73.9.2987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eckstein F., Goody R. S. Synthesis and properties of diastereoisomers of adenosine 5'-(O-1-thiotriphosphate) and adenosine 5'-(O-2-thiotriphosphate). Biochemistry. 1976 Apr 20;15(8):1685–1691. doi: 10.1021/bi00653a015. [DOI] [PubMed] [Google Scholar]
- Eckstein F. Investigation of enzyme mechanisms with nucleoside phosphorothioates. Angew Chem Int Ed Engl. 1975 Mar;14(3):160–166. doi: 10.1002/anie.197501601. [DOI] [PubMed] [Google Scholar]
- Eckstein F. Nucleoside phosphorothioates. J Am Chem Soc. 1970 Jul 29;92(15):4718–4723. doi: 10.1021/ja00718a039. [DOI] [PubMed] [Google Scholar]
- Eckstein F., Sternbach H., von der Haar F. Stereochemistry of internucleotidic bond formation by tRNA nucleotidyltransferase from baker's yeast. Biochemistry. 1977 Jul 26;16(15):3429–3432. doi: 10.1021/bi00634a021. [DOI] [PubMed] [Google Scholar]
- Hanson K. R. Concepts and perspectives in enzyme stereochemistry. Annu Rev Biochem. 1976;45:307–330. doi: 10.1146/annurev.bi.45.070176.001515. [DOI] [PubMed] [Google Scholar]
- PFLEIDERER G., ORTANDERL F. IDENTITAET VON PHOSPHODIESTERASE UND ATP-PYROPHOSPHATASE AUS SCHLANGENGIFT. Biochem Z. 1963 Jul 26;337:431–435. [PubMed] [Google Scholar]
- Saenger W., Suck D., Eckstein F. On the mechanism of ribonuclease A. Crystal and molecular structure of uridine 3'-O-thiophosphate methyl ester triethylammonium salt. Eur J Biochem. 1974 Aug 1;46(3):559–567. doi: 10.1111/j.1432-1033.1974.tb03650.x. [DOI] [PubMed] [Google Scholar]
- Usher D. A., Erenrich E. S., Eckstein F. Geometry of the first step in the action of ribonuclease-A (in-line geometry-uridine2',3'-cyclic thiophosphate- 31 P NMR). Proc Natl Acad Sci U S A. 1972 Jan;69(1):115–118. doi: 10.1073/pnas.69.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Usher D. A., Richardson D. I., Jr, Eckstein F. Absolute stereochemistry of the second step of ribonuclease action. Nature. 1970 Nov 14;228(5272):663–665. doi: 10.1038/228663a0. [DOI] [PubMed] [Google Scholar]