A compact monomeric intermediate identified by NMR in the denaturation of dimeric triose phosphate isomerase

Morgan, Charles G.; Wilkins, Deborah K.; Smith, Lorna J.; Kawata, Yasushi; Dobson, Christopher M.

doi:10.1006/jmbi.2000.3834

Cited by 37 publications

(52 citation statements)

References 27 publications

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“…All these observations suggest that in TcTIM denaturation this intermediate is an inactive expanded monomer. The presence of monomeric intermediates has been previously observed in the denaturation pattern of the yeast enzyme (yTIM) (31)(32)(33) and of the wild-type TbTIM (35).…”

Section: Discussionmentioning

confidence: 85%

“…The TIM unfolding pathways induced by guanidinium hydrochloride (Gdn-HCl) from Bacillus stearothermophilus (25), Thermotoga maritima (26), rabbit (27)(28)(29), Plasmodium falciparum (30), Saccharomyces cereVisiae (31)(32)(33), Leishmania mexicana (34), and Trypanosoma brucei (TbTIM) (35) have been studied in detail. Although the equilibrium unfolding pathways of homologous TIMs in Gdn-HCl are different, the crystallographic threedimensional structures of these enzymes are highly similar.…”

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confidence: 99%

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Reversible Equilibrium Unfolding of Triosephosphate Isomerase from Trypanosoma cruzi in Guanidinium Hydrochloride Involves Stable Dimeric and Monomeric Intermediates

et al. 2005

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The reversible guanidinium hydrochloride-induced unfolding of Trypanosoma cruzi triosephosphate isomerase (TcTIM) was characterized under equilibrium conditions. The catalytic activity was followed as a native homodimeric functional probe. Circular dichroism, intrinsic fluorescence, and size-exclusion chromatography were used as secondary, tertiary, and quaternary structural probes, respectively. The change in ANS fluorescence intensity with increasing denaturant concentrations was also determined. The results show that two stable intermediates exist in the transition from the homodimeric native enzyme to the unfolded monomers: one (N(2*)) is a slightly more expanded, non-native, and active dimer, and the other is a partially expanded monomer (M) that binds ANS. Spectroscopic and activity data were used to reach a thermodynamic characterization. The results indicate that the Gibbs free energies for the partial reactions are 4.5 (N(2) <==> N(2*)), 65.8 (N(2*) <==> 2M), and 17.8 kJ/mol (M <==> U). It appears that TcTIM monomers are more stable than those found for other TIM species (except yeast TIM), where monomer stability is only marginal. These results are compared with those for the guanidinium hydrochloride-induced denaturation of TIM from different species, where despite the functional and three-dimensional similarities, a remarkable heterogeneity exists in the unfolding pathways.

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Section: Discussionmentioning

confidence: 85%

mentioning

confidence: 99%

Reversible Equilibrium Unfolding of Triosephosphate Isomerase from Trypanosoma cruzi in Guanidinium Hydrochloride Involves Stable Dimeric and Monomeric Intermediates

et al. 2005

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“…15 However, evidence for intermediates was found in the unfolding of Plasmodium falciparum TIM 6 and Saccharomyces cerevisiae TIM. 16 Two intermediates were found in unfolding experiments of Trypanosoma brucei TIM, but unfolding of Triosephosphate Isomerase Folding this form of TIM was generally not reversible. 17 Results of an early study on the folding of rabbit muscle TIM showed that reactivation of the enzyme is second-order at low protein concentration and first-order at higher concentrations.…”

Section: Partially Folded Intermediates In Tim Folding/ Unfoldingmentioning

confidence: 98%

Equilibrium and Kinetic Folding of Rabbit Muscle Triosephosphate Isomerase by Hydrogen Exchange Mass Spectrometry

Pan

Raza

Smith

2004

Journal of Molecular Biology

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“…The first report of this monomeric intermediate was obtained at the equilibrium denaturation of yTIM in Gdn-HCl. This conformer appears at mild denaturant concentrations (41,42,43). The presence of this intermediate was also observed in the partial denaturation with the same denaturant of TbTIM (32) and in TcTIM (3) ( Table 2).…”

Section: Introductionmentioning

confidence: 59%

“…Sometimes the denaturation reaction is reversible, while in others, unspecific irreversible aggregation appears. For some species, the unfolding model is described by a two-state transition (24,(35)(36)(37)(38), while in others, more complex processes, including intermediates, have been observed (3,32,(39)(40)(41)(42)(43). These intermediates are responsible for the aggregation and irreversibility observed during unfolding experiments in vitro.…”

Section: Introductionmentioning

confidence: 98%

The Oligomeric Nature of Triosephosphate Isomerase. Studies of Monomerization

Zárate‐Pérez¹,

Vázquez‐Contreras²

2008

AIP Conference Proceedings

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A compact monomeric intermediate identified by NMR in the denaturation of dimeric triose phosphate isomerase

Cited by 37 publications

References 27 publications

Reversible Equilibrium Unfolding of Triosephosphate Isomerase from Trypanosoma cruzi in Guanidinium Hydrochloride Involves Stable Dimeric and Monomeric Intermediates

Reversible Equilibrium Unfolding of Triosephosphate Isomerase from Trypanosoma cruzi in Guanidinium Hydrochloride Involves Stable Dimeric and Monomeric Intermediates

Equilibrium and Kinetic Folding of Rabbit Muscle Triosephosphate Isomerase by Hydrogen Exchange Mass Spectrometry

The Oligomeric Nature of Triosephosphate Isomerase. Studies of Monomerization

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