The fluorescence intensity of DNA-intercalated ethidium with [ethidium]/[DNA base] being 0.005 was quenched upon the binding of another intercalating ligand, meso-tetrakis(N-methylpyridinium-4-yl)porphyrin (TMPyP). Addition of Ca 2+ enhanced the quenching efficiency. The range of separations between donor and acceptor molecules, within which total quenching occurs, was calculated using a one-dimensional resonance energy transfer mechanism to be 9.5 base-pairs or 32.3 Å in the absence of Ca 2+ ions. The distance increased to 18.7 base-pairs or about 63.6 Å in the presence 100 µM Ca
2+. Considering that (1) Ca 2+ had little effect on the binding modes of ethidium and TMPyP, which was investigated by reduced linear dichroism and (2) spectral overlap between the emission spectrum of ethidium and the absorption spectrum of TMPyP was maintained in the presence of Ca
2+, contributions from orientation factor and spectral overlap to Ca
2+-induced enhancement in DNA mediated energy transfer was limited. Although there is no direct evidence, electron transfer along the DNA stem may accompany the observed fluorescence quenching. In this respect, DNA bound Ca 2+ act as a partially conducting medium.