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J Biol Chem, Vol. 275, Issue 14, 10150-10153, April 7, 2000
,
From the Department of Chemistry-Biology, University of
Québec at Trois-Rivières, C.P. 500, Trois-Rivières, Québec G9A 5H7, Canada
Chromium(VI) salts are well known to be mutagens
and carcinogens and to easily cross the cell membranes. Because they
are powerful oxidizing agents, Cr(VI) reacts with intracellular
materials to reduce to trivalent form, which binds DNA. This study was
designed to investigate the interaction of calf thymus DNA with Cr(VI) and Cr(III) in aqueous solution at pH 6.5-7.5, using Cr(VI)/DNA(P) molar ratios (r) of 1:20 to 2:1 and Cr(III)/DNA(P)
molar ratios (r) of 1:80 to 1:2. UV-visible and Fourier transform
infrared (FTIR) difference spectroscopic methods were used to determine the metal ion-binding sites, binding constants, and the effect of
cation complexation on DNA secondary structure. Spectroscopic results
showed no interaction of Cr(VI) with DNA at low anion concentrations
(r = 1:20 to 1:1), whereas some perturbations of DNA
bases and backbone phosphate were observed at very high Cr(VI) contents
(r > 1) with overall binding constant of
K = 508 M
1. Cr(III) chelates
DNA via guanine N-7 and the nearest PO2 group with overall
binding constant of K = 3.15 × 103
M1. Evidence for cation chelate formation
comes from major shiftings and intensity variations of the guanine band
at 1717 and the phosphate asymmetric stretching vibration at 1222 cm1. At low Cr(III) concentration (r = 1:40), the number of Cr(III) ions bound to DNA were 6-7 cations/500
base pairs, and this increased to 30-35 cations/500 base pairs at high
metal ion content (r = 1:4). DNA condensation occurred
at high cation concentration (r = 1:10). No major
alteration of DNA conformation was observed, and the biopolymer
remained in the B family structure upon chromium complexation.
On sabbatical leave from Hokkaido Institute of Technology,
Sapporo, Japan.
§
To whom correspondence should be addressed. Tel.: 819-376-5052, Ext. 3310; Fax: 819-376-5084; E-mail: tajmirri@uqtr.uquebec.ca.
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