Endonuclease-like activity of heme proteins.
Heme proteins, metmyoglobin, methemoglobin, and metcytochrome c showed unusual affinity for double-stranded DNA. Calorimetric studies show that binding of methemoglobin to calf thymus DNA (CTDNA) is weakly endothermic, and the binding constant is 4.9+/-0.7x10(5) M(-1). The Soret absorption bands of the heme proteins remained unchanged, in the presence of excess CTDNA, but a new circular dichroic band appeared at 210 nm. Helix melting studies indicated that the protein-DNA mixture denatures at a lower temperature than the individual components. Thermograms obtained by differential scanning calorimetry of the mixture indicated two distinct transitions, which are comparable to the thermograms obtained for individual components, but there was a reduction in the excess heat capacity. Activation of heme proteins by hydrogen peroxide resulted in the formation of high valent Fe(IV) oxo intermediates, and CTDNA reacted rapidly under these conditions. The rate was first-order in DNA concentration, and this reactivity resulted in DNA strand cleavage. Upon activation with hydrogen peroxide, for example, the heme proteins converted the supercoiled pUC18 DNA into nicked circular and linear DNA. No reaction occurred in the absence of the heme protein, or hydrogen peroxide. These data clearly indicate a novel property of several heme proteins, and this is first report of the endonuclease-like activity of the heme proteins.
Tan
Willy B
WB
Department of Chemistry, University of Connecticut, Storrs, CT 06269-3060, USA.
Cheng
Wunhuey
W
Webber
Andrew
A
Bhambhani
Akhilesh
A
Duff
Michael R
MR
Kumar
Challa V
CV
McLendon
George L
GL
D000818:Animals; D001665:Binding Sites; D001798:Blood Proteins / Q000737:chemistry; D002151:Calorimetry; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D002942:Circular Dichroism; D045304:Cytochromes c / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004720:Endonucleases / Q000737:chemistry*; D006418:Heme / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry; D006736:Horses; D007700:Kinetics; D008786:Metmyoglobin / Q000737:chemistry; D009690:Nucleic Acid Conformation; D010084:Oxidation-Reduction; D011485:Protein Binding; D011489:Protein Denaturation; D013056:Spectrophotometry, Ultraviolet
D001798:Blood Proteins; D006454:Hemoglobins; D008786:Metmyoglobin; D006418:Heme; D045304:Cytochromes c; D004247:DNA; D004720:Endonucleases