3′-Phosphatase Activity in T4 Polynucleotide Kinase

Vicki Cameron*, Olke C. Uhlenbeck

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

221 Scopus citations

Abstract

The purification of T4 polynucleotide kinase results in the copurification of an activity which′will specifically remove the 3′-terminal phosphate from a variety of deoxyribonucleotides and ribonucleotides in the absence of ATP. This phosphatase activity requires magnesium, has a pH optimum of 6.0, and is more active with deoxyribonucleotides than ribonucleotides. T4 polynucleotide kinase and the 3′-phosphatase activity copurify by gradient elution column chromatography on DEAE-cellulose, phosphocellulose, and hydroxylapatite. The two activities are included in and comigrate on Sephadex G-200. Polyacrylamide gel electrophoresis at pH 9.2 results in comigration of the two activities together with the major protein band. The two activities respond in parallel to heat inactivation at 35 °C and ATP, a substrate for the kinase only, protects both activities from heat inactivation. It is therefore suggested that the two activities are functions of the same protein molecule.

Original languageEnglish (US)
Pages (from-to)5120-5126
Number of pages7
JournalBiochemistry
Volume16
Issue number23
DOIs
StatePublished - Nov 1 1977

ASJC Scopus subject areas

  • Biochemistry

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