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Nucleic Acids Research, 1991, Vol. 19, No. 7 1483-1489
© 1991

MOLECULAR BIOLOGY

The reaction of platinum(II) complexes with DNA. Kinetics of intrastrand crosslink formation in vitro

Frithjof Bernges and Eggehard Holler*

Institut für Biophysik und Physikalische Biochemie, Universitätsstraße 31 D-8400 Regensburg, FRG

* To whom correspondence should be addressed

Received January 9, 1991. Revised March 5, 1991. Accepted March 5, 1991.

The kinetics of the formation of bifunctional DNA platinum(II) adducts (DNA-crosslinks) have been investigated by endonuclease digestion and subsequent HPLC analysis of the soluble nucleotides and nucleotide platinum(II) adducts. The results indicate two waves of crossllnking [rate constants (0.2–0.3) min–1 and (0.015–0.025) min–1] that correlate with changes in ultra violet absorbance and ethidium bromide dependent fluorescence intensity, previously interpreted in terms of two consecutive, local conformational rearrangements of platinum-DNA (Schaller, W., Reisner, H., and Holler, E. (1987) Biochemistry 26, 943–950). The formation of crosslinks at sequences d(GpG) and d(GpNpG) follows identical kinetics. A minimal reaction mechanism is proposed for the binding of cis-diamminedichloro-platinum(II) to ONA under in vitro conditions. The approximately 3-fold higher rate for meso-[1,2-bis(2,6-dichloro-4-hydroxyphenyl)ethylenedlamine]diaqua-platinum(II) in comparison to the rate for cis-diammine-diaquaplatinum(II) indicates that crosslink formation is affected by the nature of the non-leaving platinum ligand(s).


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