A MOUSE DNA-REPAIR ENZYME (APEX NUCLEASE) HAVING EXONUCLEASE AND APURINIC APYRIMIDINIC ENDONUCLEASE ACTIVITIES - PURIFICATION AND CHARACTERIZATION

被引：81

作者：

SEKI, S

IKEDA, S

WATANABE, S

HATSUSHIKA, M

TSUTSUI, K

AKIYAMA, K

ZHANG, B

机构：

[1] Department of Molecular Biology, Institute of Cellular and Molecular Biology, Okayama University Medical School, Okayama

来源：

BIOCHIMICA ET BIOPHYSICA ACTA | 1991年 / 1079卷 / 01期

关键词：

DNA REPAIR ENZYME; EXONUCLEASE; APURINIC APYRIMIDINIC ENDONUCLEASE; DNA PHOSPHATASE; AMINO ACID SEQUENCE; PRIMING ENZYME;

D O I：

10.1016/0167-4838(91)90024-T

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A mouse repair enzyme having priming activity on bleomycin-damaged DNA for DNA polymerase was purified to apparent homogeneity and characterized. The enzyme extracted from permeabilized mouse ascites sarcoma (SR-C3H / He) cells with 0.2 M potassium phosphate buffer (pH 7.5) was purified by successive chromatographies on phosphocellulose, DEAE-cellulose, phosphocellulose (a second time), Sephadex G-100, single-stranded DNA cellulose and hydroxyapatite. The purified enzyme has an M(r) of 34000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Enzymatical studies indicated that it is a multifunctional enzyme having exonuclease, apurinic/apyrimidinic endonuclease and phosphatase activities, similar to Escherichia coli exonuclease III. This enzyme is tentatively designated as APEX nuclease for apurinic/apyrimidinic endonuclease and exonuclease activities. The amino acid composition, amino-terminal amino acid sequence and an internal amino acid sequence of APEX nuclease are determined.

引用

页码：57 / 64

页数：8

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