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PMID: 2441359 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Purification and amino-terminal amino acid sequence of an apurinic/apyrimidinic endonuclease from calf thymus.

Nucleic acids research ·Vol. 15 ·No. 14 ·1987-07-24 ·Pages 5529-44

Henner WD, Kiker NP, Jorgensen TJ, Munck JN

Abstract

An apurinic/apyrimidinic (AP) endonuclease (E.C.3.1.25.2) has been purified 1100 fold to apparent homogeneity from calf thymus by a series of ion exchange, gel filtration and hydrophobic interaction chromatographies. The purified AP endonuclease is a monomeric protein with an apparent molecular weight on SDS-PAGE of 37,000. On gel filtration the protein behaves as a protein of apparent molecular weight 40,000. DNA cleavage by this AP endonuclease is dependent on the presence of AP sites in the DNA. DNA cleavage requires the divalent cation Mg2+ and has a broad pH optimum of 7.5-9.0. Maximal rates of catalysis occur at NaCl or KCl concentrations of 25-50 mM. The amino acid composition and the amino-terminal amino acid sequence for this AP endonuclease are presented. Comparison of the properties of this AP endonuclease purified from calf thymus with the reported properties of the human AP endonuclease purified from HeLa cells or placenta indicate that the properties of such an AP endonuclease are highly conserved in these two mammalian species.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cattle Chromatography, Gel Chromatography, Ion Exchange DNA-(Apurinic or Apyrimidinic Site) Lyase Deoxyribonuclease IV (Phage T4-Induced) Endodeoxyribonucleases/isolation & purification,metabolism Kinetics Molecular Weight Substrate Specificity Thymus Gland/enzymology
Chemicals
Endodeoxyribonucleases Deoxyribonuclease IV (Phage T4-Induced) DNA-(Apurinic or Apyrimidinic Site) Lyase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Henner W D
Kiker N P
Jorgensen T J
Munck J N
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.14.5529
Published
1987-07-24
Pages
5529-44
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306005
Subset
IM
Grants
NCI NIH HHS · CA07707 · United States
NCI NIH HHS · CA35767 · United States
External Links
PubMed source DOI: 10.1093/nar/15.14.5529 PMC Full Text View journal details
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