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Nucleic Acids Research, 2001, Vol. 29, No. 13 2795-2801
© 2001 Oxford University Press

Hexamminecobalt(III)-induced condensation of calf thymus DNA: circular dichroism and hydration measurements

Besik I. Kankia*, Vitaly Buckin1 and Victor A. Bloomfield2 Department of Pharmaceutical Sciences, University of Nebraska Medical Center 986025, Omaha, NE 68198-6025, USA, 1Department of Chemistry, University College Dublin, Belfield, Dublin 4, Ireland and 2Department of Biochemistry, University of Minnesota, St Paul, MN 55108, USA

The interaction of hexamminecobalt(III), Co(NH3)63+, with 160 and 3000–8000 bp length calf thymus DNA has been investigated by circular dichroism, acoustic and densimetric techniques. The acoustic titration curves of 160 bp DNA revealed three stages of interaction: (i) Co(NH3)63+ binding up to the molar ratio [Co(NH3)63+]/[P] = 0.25, prior to DNA condensation; (ii) a condensation process between [Co(NH3)63+]/[P] = 0.25 and 0.30; and (iii) precipitation after [Co(NH3)63+]/[P] = 0.3. In the case of 3000–8000 bp DNA only two processes were observed: (i) binding up to [Co(NH3)63+]/[P] = 0.3; and (ii) precipitation after this point. In agreement with earlier observations, long DNA aggregates without changes in its B-form circular dichroism spectrum, while short DNA demonstrates a positive B->{Psi} transition after [Co(NH3)63+]/[P] = 0.25. From ultrasonic and densimetric measurements the effects of Co(NH3)63+ binding on volume and compressibility have been obtained. The binding of Co(NH3)63+ to both short and long DNA is characterized by similar changes in volume and compressibility calculated per mole Co(NH3)63+: {Delta}V = 9 cm3 mol–1 and {Delta}{kappa} = 33 x 10–4 cm3 mol–1 bar–1. The positive sign of the parameters indicates dehydration, i.e. water release from Co(NH3)63+ and the atomic groups of DNA. This extent of water displacement would be consistent with the formation of two direct, hydrogen bonded contacts between the cation and the phosphates of DNA.

* To whom correspondence should be addressed. Tel: +1 402 559 5384; Fax: +1 402 559 9543; Email: bkankia{at}unmc.edu


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