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. 1974 Dec;1(12):1763–1773. doi: 10.1093/nar/1.12.1763

Purification of polynucleotide phosphorylase by affinity chromatography and some properties of the purified enzymes

John C Smith 1, Michael AW Eaton 1
PMCID: PMC343455  PMID: 4615305

Abstract

A method is described for the preparation of p-aminophenyl oligo(dT)-Sepharose. This matrix has been used for the purification of polynucleotide phosphorylase from both E.coli and B.stearothermophilus. The effects of temperature and pH on the binding of the different enzymes to the matrix have been investigated. B.stearothermophilus isolated by affinity chromatography may be useful in selectively removing the polyA tract on the 3′-end of mRNA's.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkinson A., Jack G. W. Precipitation of nucleic acids with polyethyleneimine and the chromatography of nucleic acids and proteins on immobilised polyethyleneimine. Biochim Biophys Acta. 1973 Apr 21;308(7):41–52. doi: 10.1016/0005-2787(73)90120-2. [DOI] [PubMed] [Google Scholar]
  2. Chou J. Y., Singer M. F. Deoxyadenosine diphosphate as a substrate and inhibitor of polynucleotide phosphorylase of Micrococcus luteus. I. Deoxyadenosine diphosphate as a substrate for polymerization and the exchange reaction with inorganic 32 P. J Biol Chem. 1971 Dec 25;246(24):7486–7496. [PubMed] [Google Scholar]
  3. Chou J. Y., Singer M. F. Kinetic analysis of the phosphorolysis of oligonucleotides by polynucleotide phosphorylase. J Biol Chem. 1970 Mar 10;245(5):995–1004. [PubMed] [Google Scholar]
  4. Cuatrecasas P. Affinity chromatography. Annu Rev Biochem. 1971;40:259–278. doi: 10.1146/annurev.bi.40.070171.001355. [DOI] [PubMed] [Google Scholar]
  5. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  6. Eaton M. A., Hutchinson D. W. Poly(5-chlorocytidylic acid). Biochemistry. 1972 Aug 15;11(17):3162–3167. doi: 10.1021/bi00767a004. [DOI] [PubMed] [Google Scholar]
  7. Godefroy T. Kinetics of polymerization and phosphorolysis reactions of Escherichia coli polynucleotide phosphorylase. Evidence for multiple binding of polynucleotide in phosphorolysis. Eur J Biochem. 1970 Jun;14(2):222–231. doi: 10.1111/j.1432-1033.1970.tb00281.x. [DOI] [PubMed] [Google Scholar]
  8. Hachimori A., Muramatsu N., Noso Y. Studies on an ATPase of thermophilic bacteria. I. Purification and properties. Biochim Biophys Acta. 1970 Jun 10;206(3):426–437. doi: 10.1016/0005-2744(70)90158-0. [DOI] [PubMed] [Google Scholar]
  9. Kimhi Y., Littauer U. Z. Purification and properties of polynucleotide phosphorylase from Escherichia coli. J Biol Chem. 1968 Jan 25;243(2):231–240. [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Weatherford S. C., Weisberg L. S., Achord D. T., Apirion D. Separation of Escherichia coli ribonucleases on a DNA agarose column and the identification of an RNase H activity. Biochem Biophys Res Commun. 1972 Dec 4;49(5):1307–1315. doi: 10.1016/0006-291x(72)90609-2. [DOI] [PubMed] [Google Scholar]

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