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Thermal-unfolding Reaction of Triosephosphate Isomerase from Trypanosoma cruzi

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Abstract

Thermal denaturation of triosephosphate isomerase from Trypanosoma cruzi was studied by circular dicrhoism and fluorescence spectroscopies. The unfolding transition was found to be highly irreversible even at the very early stages of the reaction. Kinetic studies, allowed us to identify consecutive reactions. Firstly, only the tryptophan environment is altered. Next, changes on the secondary structure and hydrophobic surface exposure measured by 1-anilino-8-naphthalenesulfonate (ANS) binding were observed. Further conformational changes imply additional modifications on the secondary and tertiary structures and release of the hydrophobic dye leading to the formation of the unfolded state that is prone to aggregate.

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Abbreviations

TIM:

Triosephosphate isomerase

TcTIM:

Triosephosphate isomerase from Trypanosoma cruzi

TbTIM:

Triosephosphate isomerase from Trypanosoma brucei

yTIM:

Yeast triosephosphate isomerase

CD:

Circular dichroism

Tris:

Tris(hydroxy-methyl) aminomethane

Gdn-HCl:

Guanidine hydrochloride

TEA:

Triethanolamine

SCM:

Spectral centre of mass

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Acknowledgements

We are very grateful to Dr. Armando Gomez Puyou and Beatriz Aguirre for the facilities and help given throughout the purification of TcTIM. EMH received financial support from Instituto Politécnico Nacional and PIFI-IPN (20050356 and 20060916), LMMV thanks economic sustain from CONACyT (175886). This work was supported by grants from TWAS (04–352 RG/BIO/LA), CONACyT (45990 and 46168-M), ECOS m05–501 and SIP-IPN 20070141.

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Authors and Affiliations

  1. Laboratorio de Investigación Bioquímica, Programa Institucional en Biomedicina Molecular ENMyH-IPN, Guillermo Massieu Helguera No. 239, La Escalera Ticoman, 07320, D.F, Mexico

    Edgar Mixcoha-Hernández & Claudia G. Benítez-Cardoza

  2. Área de Biofisicoquímica, Departamento de Química, Universidad Autónoma Metropolitana-Iztapalapa, Apartado Postal 55–534, Iztapalapa, 09340, D.F, Mexico

    Liliana M. Moreno-Vargas & Arturo Rojo-Domínguez

  3. Departamento de Ciencias Naturales, Universidad Autónoma Metropolitana-Cuajimalpa, Pedro Antonio de los Santos 84, San Miguel Chapultepec, 11850, D.F, Mexico

    Arturo Rojo-Domínguez

Authors
  1. Edgar Mixcoha-Hernández
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  2. Liliana M. Moreno-Vargas
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  3. Arturo Rojo-Domínguez
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  4. Claudia G. Benítez-Cardoza
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Corresponding author

Correspondence to Claudia G. Benítez-Cardoza.

Additional information

Edgar Mixcoha-Hernández and Liliana M. Moreno-Vargas contributed equally to this work

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Mixcoha-Hernández, E., Moreno-Vargas, L.M., Rojo-Domínguez, A. et al. Thermal-unfolding Reaction of Triosephosphate Isomerase from Trypanosoma cruzi . Protein J 26, 491–498 (2007). https://doi.org/10.1007/s10930-007-9090-3

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