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Denaturation Studies by Fluorescence and Quenching of Thermophilic Protein NAD+-Glutamate Dehydrogenase from Thermus thermophilus HB8

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Abstract

Fluorescence techniques have been used to study the structural characteristics of many proteins. The thermophilic enzyme NAD-glutamate dehydrogenase from Thermus thermophilus HB8 is found to be a hexameric enzyme. Fluorescence spectra of native and denatured protein and effect of denaturants as urea and guanidine hydrochloride on enzyme activity of thermophilic glutamate dehydrogenase (t-GDH) have been analyzed. Native t-GDH presents the maximum emission at 338 nm. The denaturation process is accompanied by an exposure to the solvent of the tryptophan residues, as manifested by the red shift of the emission maximum. Fluorescence quenching by external quenchers, KI and acrylamide, has also been carried out.

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Authors and Affiliations

  1. División de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Alicante, Alicante, Spain

    Jose L. Ruiz, Juan Ferrer, Carmen Pire, Francisco I. Llorca & Maria José Bonete

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  1. Jose L. Ruiz
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  2. Juan Ferrer
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  3. Carmen Pire
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  4. Francisco I. Llorca
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  5. Maria José Bonete
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Correspondence to Maria José Bonete.

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Ruiz, J.L., Ferrer, J., Pire, C. et al. Denaturation Studies by Fluorescence and Quenching of Thermophilic Protein NAD+-Glutamate Dehydrogenase from Thermus thermophilus HB8. J Protein Chem 22, 295–301 (2003). https://doi.org/10.1023/A:1025080722424

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  • DOI: https://doi.org/10.1023/A:1025080722424

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