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Anion concentration modulates the conformation and stability of the molten globule of cytochrome c

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Abstract

Anions induce collapse of acid-denatured cytochrome c into a compact state, the A-state, showing molten globule character. Since structural information on partially folded forms of proteins is important for a deeper understanding of folding mechanisms and of the factors affecting protein stabilization, in this paper we have investigated in detail the effects of anions on the tertiary conformation of the A-state. We have found that the salt-induced collapse of acid-denatured cytochrome c leads to a number of equilibria between high-spin and low-spin heme states and between two types of low-spin states. The two latter states are characterized by conformations leading to a native-like Met-Fe-His axial coordination and a bis-His configuration. The equilibrium between these two A-states is dependent on the concentration and/or size of the anions (i.e. the bigger the anion, the greater its effect). Further, on the basis of fast kinetic data, a kinetic model of the folding process from the acid-unfolded protein to the A-state (at low and high anion concentration) is described.

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Abbreviations

cyt c :

cytochrome c

cyt c′′:

cytochrome c′′

LS:

low-spin

HS:

high-spin

RR:

resonance Raman

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Acknowledgements

This research was funded in part by grants from MIUR (COFIN 2001031798).

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Authors and Affiliations

  1. Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università di Roma "Tor Vergata", V. Montpellier 1, 00133 , Rome, Italy

    Federica Sinibaldi, Chiara Ciaccio, Massimo Coletta & Roberto Santucci

  2. Dipartimento di Chimica, Università degli Studi di Firenze, V. della Lastruccia 3, 50019 , Sesto Fiorentino, Italy

    Barry D. Howes & Giulietta Smulevich

Authors
  1. Federica Sinibaldi
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  2. Barry D. Howes
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  3. Giulietta Smulevich
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  4. Chiara Ciaccio
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  5. Massimo Coletta
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  6. Roberto Santucci
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Correspondence to Roberto Santucci.

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Sinibaldi, F., Howes, B.D., Smulevich, G. et al. Anion concentration modulates the conformation and stability of the molten globule of cytochrome c . J Biol Inorg Chem 8, 663–670 (2003). https://doi.org/10.1007/s00775-003-0462-7

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  • DOI: https://doi.org/10.1007/s00775-003-0462-7

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