期刊名称:Proceedings of the National Academy of Sciences
印刷版ISSN:0027-8424
电子版ISSN:1091-6490
出版年度:1980
卷号:77
期号:6
页码:3201-3205
DOI:10.1073/pnas.77.6.3201
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:Using a filter binding assay, we have detected and partially purified a protein from human placenta that has a high affinity for N-acetoxy-2-acetylaminofluorene-modified double-stranded DNA (AAF-[3H]DNA) of bacteriophage T7. This protein has been partially purified from a 1 M NaCl extract of a crude nuclear fraction by a combination of ion-exchange and nucleic acid affinity chromatography. With AAF-[3H]DNA as the substrate, the binding reaction reached equlibrium within 1 hr at 4 degrees C, and the extent of binding ws proportional to the amount of protein added. Complex formation was dependent on both pH and salt concentration and was unaffected by the presence of sulfhydryl-blocking agents. The purest protein fraction also recognizes DNA modified with methylmethane-sulfonate or methylnitrosourea. It shows little or no recognition of single-stranded DNA, double-stranded DNA, supercoiled bacteriophage phiX174 DNA, partially depurinated DNA, glucosylated bacteriophage T4DNA, or UV-irradiated DNA. No endo- or exonuclease activity, DNA polymerase activity, or glucosylase activity for AAF-DNA was detectable in the preparation.