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Chemical Unfolding of Enolase from Saccharomyces cerevisiae Exhibits a Three-State Model

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Abstract

Enolase is a multifunctional protein that participates in glycolysis and gluconeogenesis and can act as a plasminogen receptor on the cell surface of several organisms, among other functions. Despite its participation in a variety of biological and pathophysiological processes, its stability and folding/unfolding reaction have not been fully explored. In this paper we present, the urea and GdnHCl-induced denaturation of enolase studied by means of fluorescence and circular dichroism spectroscopies. We found that enolase unfolds through a highly reversible pathway, populating a stable intermediate species in a range of experimental conditions. The refolding reaction also exhibits an intermediate state that might have a slightly more compact conformation compared to the unfolding intermediate. The thermodynamic parameters associated with the unfolding reaction are presented and discussed.

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Abbreviations

TRIS:

TRIS(hydroxy-methyl) aminomethane

GdnHCl:

Guanidinium hydrochloride

SCM:

Spectral center of mass

CD:

Circular dichroism

SASA:

Solvent-accessible surface area

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Acknowledgments

We are very thankful with Dr. Andrés Hernández Arana for the use of the circular dichroism spectrometer. NCI was supported by a fellowship from CONACyT (204849) and from the Instituto Politécnico Nacional with a PIFI-IPN student grant. This work was supported by grants from TWAS (04-352 RG/BIO/LA), CONACyT (45990), and SIP-IPN (20080356, 20091074).

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Authors and Affiliations

  1. Laboratorio de Investigación Bioquímica, Programa Institucional en Biomedicina Molecular, Sección de Estudios de Posgrado, ENMyH-Instituto Politécnico Nacional, Guillermo Massieu Helguera No. 239, La Escalera Ticoman, DF, 07320, Mexico

    Dénison S. Sánchez-Miguel, Jahir Romero-Jiménez, César A. Reyes-López, Ana Lilia Cabrera-Ávila, Normande Carrillo-Ibarra & Claudia G. Benítez-Cardoza

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  1. Dénison S. Sánchez-Miguel
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  2. Jahir Romero-Jiménez
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  3. César A. Reyes-López
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  4. Ana Lilia Cabrera-Ávila
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  5. Normande Carrillo-Ibarra
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  6. Claudia G. Benítez-Cardoza
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Correspondence to Claudia G. Benítez-Cardoza.

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Sánchez-Miguel, D.S., Romero-Jiménez, J., Reyes-López, C.A. et al. Chemical Unfolding of Enolase from Saccharomyces cerevisiae Exhibits a Three-State Model. Protein J 29, 1–10 (2010). https://doi.org/10.1007/s10930-009-9215-y

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