Thermal unfolding of apo- and holo-enolase from Saccharomyces cerevisiae: Different mechanisms, similar activation enthalpies

Moreno-Vargas, Liliana M.; Normande, Carrillo-Ibarra; Arzeta-Pino, Lilian; Benítez‐Cardoza, Claudia G.

doi:10.1016/j.ijbiomac.2011.07.021

Cited by 3 publications

(6 citation statements)

References 43 publications

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“…From the slope of this plot, the activation enthalpy (ΔH ≠ ) associated with the thermal unfolding of holo-HpEno was calculated (Table 2). In the same table, the corresponding ΔH ≠ reported for yeast holoenolase [28] is also shown. Interestingly, both activation enthalpies are equivalent.…”

Section: Resultsmentioning

confidence: 95%

“…The kinetics of the thermal denaturation reaction was also studied for holo-HpEno. The activation enthalpy associated with the unfolding process is almost equivalent between yeast enolase [28] and recombinant HpEno (obtained at similar experimental conditions). The activation enthalpy measures the disruption of enthalpically favourable interactions in the folded state that occur to achieve the transition state.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, the activation enthalpy is not calculated from the absolute value of the rate constants of denaturation but from their dependence on temperature. Therefore, two proteins with different conformational stabilities can show similar activation enthalpies for thermal unfolding [28].…”

Section: Discussionmentioning

confidence: 99%

“…Thermal denaturation transitions were carried out as described before [28–31] (also see the methodological details and data analysis in the supplementary materials: appendix 1).…”

Section: Methodsmentioning

confidence: 99%

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Biochemical and Biophysical Characterization of the Enolase fromHelicobacter pylori

López-López

Rodríguez-Luna

Lara-Ramírez

et al. 2018

BioMed Research International

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Enolase, which catalyses the conversion of 2-phospho-D-glycerate to phosphoenolpyruvate, is an important enzyme in the classic glycolysis pathway in cells. Enolase is highly conserved in organisms from bacteria to humans, indicating its importance in cells. Thus, enolase is a good target for developing new drugs. In the last decade, new functions of this enzyme have been found. Helicobacter pylori is a common human pathogen that causes gastric diseases and even gastric cancer. In this study, the sequence of H. pylori enolase (HpEno) was analysed; the conservation (at least partial) of binding sites for cofactor, plasminogen, and host extracellular RNA, as well as catalytic site, indicates that HpEno should be capable of performing the functions. Recombinant HpEno was overexpressed and purified from E. coli. Compared to the enolases from other species, HpEno had similar characteristics for its secondary structure. The temperature-induced profiles indicate that HpEno is quite stable to temperature, compared to other homologs. Regarding the kinetics of the unfolding reaction, we found that the activation enthalpy associated with the thermal unfolding reaction is equivalent to the reported activation enthalpy for yeast enolase, indicating a similar scaffold and kinetic stability. Although a wide range of experimental conditions were assayed, it was not possible to detect any enzymatic activity of HpEno. To prove the lack of activity, still a much wider range of experiments should be carried out.

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Section: Resultsmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…Thermal denaturation transitions were carried out as described before [28–31] (also see the methodological details and data analysis in the supplementary materials: appendix 1).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Biochemical and Biophysical Characterization of the Enolase fromHelicobacter pylori

López-López

Rodríguez-Luna

Lara-Ramírez

et al. 2018

BioMed Research International

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“…This means that the divalent cofactor stabilized enolase from T. vaginalis, as observed for enolase from other biological species as well as other enzymes that are stabilized by their cofactors. , In the case of TvEno, subunit interactions necessary to increase the percentage of dimers and oligomers as observed in Tris–HCl and Tris–acetate buffers upon cofactor binding might have contributed to the observed thermal stabilization (Figure ). Commonly, from thermal denaturation profiles, it is possible to obtain thermodynamic parameters associated with the unfolding or refolding reaction when a denaturation model can be assumed to describe the experimental data.…”

Section: Resultsmentioning

confidence: 75%

Self-Association of Enolase from Trichomonas vaginalis. Monomers, Dimers, and Octamers Coexist in Solution

et al. 2018

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We used small-angle X-ray scattering to study the self-association of enolase from Trichomonas vaginalis as a function of the protein concentration and cosolute type. We observed coexisting monomers, dimers, and octamers in variable relative populations, depending on whether Tris− acetate, Tris−HCl, or potassium phosphate buffers were used. Phosphate ions hindered the formation of dimers and octamers. In contrast, the populations of dimers and octamers increased in Tris−acetate or Tris−HCl buffers and additionally increased by augmenting protein concentration or adding magnesium. Single oligomeric species could not be isolated in any of the experimental conditions tested. Furthermore, the secondary and tertiary structures, as well as the temperatureinduced denaturation of the mixtures of species, were investigated. The acquired species lost enzymatic activity, but they were prone to interact with plasminogen, as judged from changes in the secondary and tertiary structures upon complex formation.

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The chaperone role of the pyridoxal 5′-phosphate and its implications for rare diseases involving B6-dependent enzymes

Cellini

Montioli

Oppici

et al. 2014

Clinical Biochemistry

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Thermal unfolding of apo- and holo-enolase from Saccharomyces cerevisiae: Different mechanisms, similar activation enthalpies

Cited by 3 publications

References 43 publications

Biochemical and Biophysical Characterization of the Enolase fromHelicobacter pylori

Biochemical and Biophysical Characterization of the Enolase fromHelicobacter pylori

Self-Association of Enolase from Trichomonas vaginalis. Monomers, Dimers, and Octamers Coexist in Solution

The chaperone role of the pyridoxal 5′-phosphate and its implications for rare diseases involving B6-dependent enzymes

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