Binding of cationic porphyrin to isolated DNA and nucleoprotein complex: quantitative analysis of binding forms under various experimental conditions.
We studied the complex formation of tetrakis(4-N-methylpyridyl)porphyrin (TMPyP) with double stranded DNAs and T7 phage nucleoprotein complex. We analyzed the effect of base pair composition of DNA, the presence of capsid protein, and the composition of the microenvironment on the distribution of TMPyP between binding forms as determined by the decomposition of porphyrin absorption spectra. No difference was found in the amount of bound TMPyP between DNAs of various base compositions; however, the ratio of TMPyP binding forms depends on the AT/GC ratio. The presence of protein capsid opposes the binding of TMPyP to DNA. This behavior offers a possibility to investigate the protein capsid integrity due to the analysis of porphyrin binding. Increasing ionic strength of monovalent ions decreases the amount of bound porphyrin through the inhibition of intercalation, but does not influence the quantity of groove-binding forms when TMPyP interacts with isolated DNA. In the case of the nucleoprotein complex the groove-binding is also inhibited already at 140 mM ionic strength. The presence of 1 mM divalent cations (Mg(2+), Ca(2+), Cu(2+) and Ni(2+)) in a buffer solution of 70 mM ionic strength does not influence significantly the free to bound ration of TMPyP when it interacts with isolated DNA. The contribution of binding forms is remarkably different in Mg(2+)/Ca(2+) and Cu(2+)/Ni(2+) containing solutions. Transition metals significantly decrease the binding sites for intercalation in both DNA and nucleoprotein complex, but facilitate the groove-binding of TMPyP to isolated DNA.
Zupán
Kristóf
K
Institute of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary.
Herényi
Levente
L
Tóth
Katalin
K
Egyeki
Marianna
M
Csík
Gabriella
G
D000818:Animals; D001482:Base Composition; D001665:Binding Sites; D002413:Cations, Divalent / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007364:Intercalating Agents / Q000737:chemistry / Q000378:metabolism*; D009698:Nucleoproteins / Q000378:metabolism*; D009994:Osmolar Concentration; D011166:Porphyrins / Q000737:chemistry / Q000378:metabolism*; D013696:Temperature; D014325:Tromethamine / Q000737:chemistry
D002413:Cations, Divalent; D007364:Intercalating Agents; D009698:Nucleoproteins; D011166:Porphyrins; D014325:Tromethamine; C021096:tetra(4-N-methylpyridyl)porphine; D004247:DNA