DNA-binding proteins of human placenta: purification and characterization of an endonuclease.
DNA binding proteins present in the cytoplasm and nuclei of term placenta were isolated by DNA-cellulose chromatography and analysed by electrophoresis in high resolution polyacrylamide gradient gels. A denatured DNA specific protein of approximate molecular weight 34 000 daltons was the predominant DNA binding protein of the cytoplasm; this protein consisted of over 65% of the total DNA binding proteins of the 0.15 M NaCl eluate of the cytoplasm. The cytoplasmic extracts contained two additional DNA binding proteins of molecular weight 24 000 and 18 000 daltons and these proteins bound preferentially to ds DNA. All the three DNA binding proteins were also present in the nuclei and electrophoresis of histones in adjacent lanes indicated that they are not histones. The 34 000-dalton DNA binding protein has been purified by ammonium sulphate fractionation followed by phosphocellulose (PC) chromatography. The DBP eluted from the PC column between 0.125-0.15 M potassium phosphate. PC fractions containing electrophoretically pure 34 KD DBP showed an endonuclease activity capable of converting plasmid pBR 322 DNA to the linear form. Maximum endonucleolytic activity was observed in the presence of 3-5 mM Mg2+ and the enzyme activity was completely inhibited by 3 mM ethylenediamine tetraacetate.
D002467:Cell Nucleus / Q000032:analysis; D003593:Cytoplasm / Q000032:analysis; D004268:DNA-Binding Proteins / Q000302:isolation & purification*; D004720:Endonucleases / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010920:Placenta / Q000032:analysis* / Q000201:enzymology; D011247:Pregnancy
D004268:DNA-Binding Proteins; D004720:Endonucleases; D008274:Magnesium