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.

Keywords

heme proteins / divalent cations / fluorescence quenching / isothermal calorimetry / time correlated single photon counting (TCSPC)

References

[1] Arias-Moreno X, Abian O, Vega S, Sancho J, Velazquez-Campoy A (2011). Protein-cation interactions: structural and thermodynamic aspects. Curr Protein Pept Sci, 12(4): 325–338

[2] Bardhan M, Mandal G, Ganguly T J (2009). Steady state, Time resolved and Circular dichroism spectroscopic studies to reveal the nature of interactions of zinc oxide nanoparticles with transport protein Bovine Serum Albumin and to monitor the possible protein conformational changes. Appl Phys (Berl), 106(3): 34701–34705

[3] Basak P, Bhattacharyya M (2013). Intrinsic tryptophan fluorescence and related energy transfer in Leghemoglobin isolated from Arachis hypogeal. Turkish. J Biochem, 38: 9–13

[4] Basak P, Pattanayak R, Bhattacharyya M (2015). Transition metal induced conformational change of heme proteins. Spectrosc Lett, 48(5): 324–330

[5] Berezin M Y, Achilefu S (2010). Fluorescence lifetime measurements and biological imaging. Chem Rev, 110(5): 2641–2684

[6] Berlett B S, Levine R L, Stadtman E R (2000). Use of isosbestic point wavelength shifts to estimate the fraction of a precursor that is converted to a given product. Anal Biochem, 287(2): 329–333

[7] Cantor C R, Schimmel P R (1984). Biophysical Chemistry Part II:Techniques for the Study of Biological Structure and Function. New York: W.H.Freeman and Company, 386–398

[8] Das P, Mallik A, Halder B, Chakraborty A, Chattopadhyay N (2006). Effect of nanocavity confinement on the rotational relaxation dynamics: 3-acetyl-4-oxo-6,7-dihydro- 12 H indolo- [ 2, 3- a ] quinolizine in micelles. J Chem Phys, 125(4): 044516

[9] Dickerson R E, Timkovich R (1975). The Enzymes, (P. Boyer Ed.). New York: Academic Press

[10] Faergeman N J, Sigurskjold B W, Kragelund B B, Andersen K V, Knudsen J (1996). Thermodynamics of ligand binding to acyl-coenzyme A binding protein studied by titration calorimetry. Biochemistry, 35(45): 14118–14126

[11] Giovannetti R, Uddin J (2012). The Use of Spectrophotometry UV-Vis for the Study of Porphyrins, InTech, 95–96, ISBN: 978–953–51–0664–7

[12] Gourion-Arsiquaud S, Chevance S, Bouyer P, Garnier L, Montillet J L, Bondon A, Berthomieu C (2005). Identification of a Cd²⁺- and Zn²⁺-binding site in cytochrome c using FTIR coupled to an ATR microdialysis setup and NMR spectroscopy. Biochemistry, 44(24): 8652–8663

[13] Harbury H A, Loach P A (1960). Oxidation-linked proton functions in heme octa- and undecapeptides from mammalian cytochrome c. J Biol Chem, 235: 3640–3645

[14] Heringa J, Argos P (1991). Side-chain clusters in protein structures and their role in protein folding. J Mol Biol, 220(1): 151–171

[15] Hua Y J, Liua Y I, Zhanga L X, Zhaoa R M, Qua S S (2005). Studies of interaction between colchicine and bovine serum albumin by fluorescence quenching method. J Mol Struct, 750(1-3): 174–178

[16] Kadish K M, Smith K M, Guilard R (2010). Handbook of Porphyrin Science. World Scientific Publishing: Singapore

[17] Kahn K, Bruice T C (2003). Comparison of reaction energetics and leaving group interactions during the enzyme-catalyzed and uncatalyzed displacement of chloride from haloalkanes. J Phys Chem B, 107(28): 6876–6685

[18] Kang J, Liu Y, Xie M X, Li S, Jiang M, Wang Y D (2004). Interactions of human serum albumin with chlorogenic acid and ferulic acid. Biochim Biophys Acta, 1674(2): 205–214

[19] Lakowicz J R (2006). Principles of Fluorescence Spectroscopy, 3rd ed. New York: Plenum Press, 277–285

[20] Lehrer S S (1971). Solute perturbation of protein fluorescence. The quenching of the tryptophyl fluorescence of model compounds and of lysozyme by iodide ion. Biochemistry, 10(17): 3254–3263

[21] Liao M S, Watts J D, Huang M J (2006). DFT/TDDFT study of lanthanide(III) mono- and bisporphyrin complexes. J Phys Chem A, 110(48): 13089–13098

[22] Marchon J C, Mashiko T, Reed C A (1982). Electron-Transport and Oxygen Utilization. (C.H.O. Ed.) North Holland, New York: Elsevier

[23] Mata L, Sanchez L, Calvo M (1997). Interaction of mercury with human and bovine milk proteins. Biosci Biotechnol Biochem, 61(10): 1641–1645

[24] Mátyuss L, Szöllosi J, Jenei A (2006). Steady-state fluorescence quenching applications for studying protein structure and dynamics. J Photochem Photobiol B, 83(3): 223–236

[25] Muller P (1994). Glossary of terms used in physical organic chemistry. Pure Appl Chem, 66(5): 1077

[26] Murayama M (1958). Titrable sulphahydryl groups of Hemoglobin C and fetal Hemoglobin at 0° and 38°. J Biol Chem, 230: 163–168

[27] Murayama M (1959). On the nature of the interaction between binding sites for heavy metals (mercapto-mercapto interactions) in normal human hemoglobin. J Biol Chem, 234: 3158–3162

[28] Murphy C B, Zhang Y, Troxler T, Ferry V, Martin J J, Jones W E (2004). Probing Förster and Dexter energy-transfer mechanisms in fluorescent conjugated polymer chemosensors. J Phys Chem, 108(5): 1537–1543

[29] Nada T, Terazima M (2003). A novel method for study of protein folding kinetics by monitoring diffusion coefficient in time domain. Biophys J, 85(3): 1876–1881

[30] Ordway G A, Garry D J (2004). Myoglobin: an essential hemoprotein in striated muscle. J Exp Biol, 207(Pt 20): 3441–3446

[31] Pauling L (1960). The Nature of the Chemical Bond (3rd Edn.), Ithaca, NY: Cornell University Press

[32] Pinto M R, Schanze K S (2004). Amplified fluorescence sensing of protease activity with conjugated polyelectrolytes. Proc Natl Acad Sci USA, 101(20): 7505–7510

[33] Raphael A L, Gray H B (1991). Semisynthesis of axial-ligand (position 80) mutants of cytochrome c. J Am Chem Soc, 113(3): 1038–1040

[34] Rispens T, Lakemond C M M, Derksen N I, Aalberse R C (2008). Detection of conformational changes in immunoglobulin G using isothermal titration calorimetry with low-molecular-weight probes. Anal Biochem, 380(2): 303–309

[35] Ross P D, Subramanian S (1981). Thermodynamics of protein association reactions: forces contributing to stability. Biochemistry, 20(11): 3096–3102

[36] Samanta U, Pal D, Chakrabarti P (2000). Environment of tryptophan side chains in proteins. Proteins, 38(3): 288–300

[37] Takeda K, Hachiya K (2002). Interaction of Protein with Ionic Surfactant, Part I: Marcel Dekker; New York

[38] Wachter R M, Elsliger M A, Kallio K, Hanson G T, Remington S J (1998). Structural basis of spectral shifts in the yellow-emission variants of green fluorescent protein. Structure, 6(10): 1267–1277

[39] Walker V E, Castillo N, Matta C F, Boyd R J. (2010). The Effect of multiplicity on the size of iron(II) and the structure of iron(ii) porphyrins. J Phys Chem A, 114:10315–10319

[40] Wang J, Guo D, Yuan X (2006). Influence of copper on the interaction between cytochrome c and sulfite in vitro. J Biochem Mol Toxicol, 20(5): 255–258

[41] Wuthrich R Q (1985). Amino acid sequence, haem iron coordination geometry and functional properties of mitochondrial and bacterial c-type cytochromes. Rev Biophys, 18(02): 111–134

[42] Yan Y, Marriott G (2003). Analysis of protein interactions using fluorescence technologies. Curr Opin Chem Biol, 7(5): 635–640

[43] Zaidi N, Ahmad E, Rehan M, Rabbani G, Ajmal M R, Zaidi Y, Subbarao N, Khan R H (2013). A comprehensive insight into binding of hippuric acid to human serum albumin: A study to uncover its impaired elimination through hemodialysis. J Phys Chem B, 117: 2595–2604

[44] Zhang Y Z, Xiang X, Mei P, Dai J, Zhang L L, Liu Y (2009). Spectroscopic studies on the interaction of Congo Red with bovine serum albumin. Spectrochim Acta A Mol Biomol Spectrosc, 72(4): 907–914

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

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg