Biochemistry (Moscow)

, Volume 80, Issue 4, pp 463–472 | Cite as

Denaturation properties and folding transition states of leghemoglobin and other heme proteins

  • Pijush Basak
  • Niloy Kundu
  • Rudradip Pattanayak
  • Maitree BhattacharyyaEmail author


This work reports unfolding transitions of monomeric heme proteins leghemoglobin (Lb), myoglobin (Mb), and cytochrome c (Cyt c) utilizing UV-Vis spectra, steady-state and time-resolved fluorescence methods. Conformational stabilities of the native “folded” state of the proteins and their “unfolded” states were investigated in the light of a two-state transition model. Two-state transition values for ΔGD (298K) were obtained by denaturation with the chaotropic agents urea and guanidium hydrochloride (GdnHCl). The free energy value of Lb is the lowest compared to Cyt c and Mb along the denaturation pathway. The m value is also the lowest for Lb compared to Cyt c and Mb. The m value (a measure of dependence of ΔGD on denaturant concentration) for Cyt c and Mb is lower when it is denatured with urea compared to GdnHCl. The UV-Vis absorbance maximum and steady state fluorescence emission maximum were drastically red shifted in the presence of a certain denaturant concentration both in cases of Mb and Lb, but the scenario is different for Cyt c. The results are analyzed using a two-state transition model. The lifetime data clearly indicate the presence of an intermediate state during denaturation. The unfolding transition can modulate the conformation, stability, and surface exposure of these biologically important proteins.

Key words

chaotropic agent denaturation heme protein Gibbs free energy two-state transition 


Cyt c

cytochrome c


guanidium hydrochloride








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Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  • Pijush Basak
    • 1
  • Niloy Kundu
    • 2
  • Rudradip Pattanayak
    • 1
  • Maitree Bhattacharyya
    • 1
    Email author
  1. 1.Department of BiochemistryUniversity of CalcuttaKolkataIndia
  2. 2.Department of ChemistryIndian Institute of TechnologyKharagpurIndia

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