Highly selective affinity labeling of the primer-binding site of E.coli DNA polymerase I

R. L. Mitina, A. A. Mustaev, E. F. Zaychikov, V. V. Khomov, O. I. Lavrik

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Highly selective affinity labeling of the primer site of E.coli DNA polymerase I was performed with the 5'-reactive derivatives of oligothymidylate in the presence of poly(dA) template. Subtilysine cleavage proved that the site of affinity modification belonged to the 'Klenow' part of DNA polymerase I. If taken separately, Klenow fragment was not labeled by these oligonucleotide derivatives. The sites of affinity labeling were tested in the structure of DNA polymerase I by hydroxylamine cleavage. At least two sites of labeling were revealed. The main one was localized between Gly-833 and His-928.

Original languageEnglish (US)
Pages (from-to)181-183
Number of pages3
JournalFEBS Letters
Volume272
Issue number1-2
DOIs
StatePublished - Oct 15 1990
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Molecular Biology
  • Biophysics
  • Structural Biology
  • Biochemistry
  • Cell Biology

Keywords

  • Affinity labeling
  • E.coli DNA polymerase I
  • Klenow fragment

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