Nonproteolyzed Form of DNA-Polymerase ε from T-Cell Spontaneous Lymphoma of Sprague-Dawley Inbred Rat: Isolation and Characterization

Autor:	Antonín Holý, Pavel Kramata, Ivan Votruba, Otová B, Miroslav Otmar, Gabriel Birkus
Rok vydání:	1998
Předmět:	chemistry.chemical_classification Exonuclease biology DNA polymerase Protein subunit General Chemistry Processivity Molecular biology Enzyme assay Enzyme chemistry Biochemistry biology.protein Nucleotide Binding site
Zdroj:	Collection of Czechoslovak Chemical Communications. 63:723-731
ISSN:	1212-6950 0010-0765
Popis:	Using a simple isolation procedure and selective assay for the determination of enzyme activity the nonproteolyzed and proteolyzed form of DNA-polymerase ε (pol ε and pol ε*) from the lymphoma of Sprague-Dawley inbred rats were purified. Nonproteolyzed pol ε is composed of two subunits (240 000 and 50 000) with sedimentation coefficient 10.5 S, while the subunit composition of pol ε* was 145 000 and 73 000. Estimated Km values for dATP and dGTP as well as Ki values for acyclic nucleotide analogs (PMEApp, HPMPApp and PMEDAPpp) in pol ε and pol ε* catalyzed reactions have shown that a proteolysis probably does not affect pol ε binding site for dNTPs. Both enzymes (pol ε and pol ε*) possess 3'-5'-exonuclease activity with different Km for 3'-OH end of template poly dA-oligo dT18 (1.6 μmol/l and 0.36 μmol/l, respectively).
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::6e3a94ba1f2c8ab05880adae9eb99b96 https://doi.org/10.1135/cccc19980723 Zobrazit plný text záznamu