Selective binding of divalent cations toward heme proteins

Abstract

Potential toxicity of transition metals like Hg, Cu and Cd are well known and their affinity toward proteins is of great concern. This work explores the selective nature of interactions of Cu²⁺, Hg²⁺ and Cd²⁺ with the heme proteins leghemoglobin, myoglobin and cytochrome C. The binding profiles were analyzed using absorbance spectrum and steady-state fluorescence spectroscopy. Thermodynamic parameters like enthalpy, entropy and free energy changes were derived by isothermal calorimetry and consequent binding parameters were compared for these heme proteins. Free energy (DG) values revealed Cu²⁺ binding toward myoglobin and leghemoglobin to be specific and facile in contrast to weak binding for Hg²⁺ or Cd²⁺ . Time correlated single photon counting indicated significant alteration in excited state lifetimes for metal complexed myoglobin and leghemoglobin suggesting bimolecular collisions to be involved. Interestingly, none of these cations showed significant affinity for cytochrome c pointing that, presence of conserved sequences or heme group is not the only criteria for cation binding toward heme proteins, but the microenvironment of the residues or a specific folding pattern may be responsible for these differential conjugation profile. Binding of these cations may modulate the conformation and functions of these biologically important proteins.

Key words： heme proteins; divalent cations; fluorescence quenching; isothermal calorimetry; time correlated single photon counting (TCSPC)

Acknowledgements

We acknowledge World Bank for providing fellowship to P. Basak. We are thankful to UGC-UPE (UGC/191/UPE/ 2007), DST (FIST), World Bank (54 ICZMP), UGC-CAS, DBT-IPLS Government of India for providing the instrumental and infrastructural facility in Department of Biochemistry, C.U.

Pijush Basak , Tanay Debnath , Rajat Banerjee and Maitree Bhattacharyya declare that they have no conflict of interest. This article does not contain any studies with human or animal subjects performed by any of the authors

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