Predissociated dimers and molten globule monomers in the equilibrium unfolding of yeast glutathione reductase
Open accessAbstract
The equilibrium unfolding of dimeric yeast glutathione reductase (GR) by guanidine hydrochloride (GdnHCl) was investigated. Unfolding was monitored by a variety of techniques, including intrinsic fluorescence emission, anisotropy and iodide quenching measurements, far-ultraviolet circular dichroism and thiol reactivity measurements. At 1 M GdnHCl, one thiol group of GR became accessible to modification with 5,5'-dithiobis-(2-nitrobenzoic) acid (DTNB), whereas no changes could be detected in the spectroscopic properties (fluorescence, circular dichroism) of the protein. Between 2 and 3 M GdnHCl, two partially folded intermediate states possessing flexible tertiary structures (revealed by fluorescence data) but compact secondary structures (as indicated by circular dichroism measurements) were identified. The quaternary structure of GR in the presence of GdnHCl was also investigated by size-exclusion liquid chromatography. These results indicated the presence of an expanded predissociated dimer at 2.5 M GdnHCl and partially folded monomers at 3 M GdnHCl. Taken together, these results suggest the existence of two molten-globule-like intermediate species (one dimeric and one monomeric) in the unfolding of GR. The results are discussed in terms of the mechanism of GR folding and dimerization.
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Variations in chronotype between urban and rural areas within the same country have been observed in Japanese adolescents and in adults in Brazil, India, and elsewhere.
“In addition to country-specific differences, variations in chronotype between urban and rural areas within the same country have been observed in Japanese adolescents (53, 59, 60) and in adults in Brazil, India, and elsewhere (61–63).”
From Chronotype: Implications for Epidemiologic Studies on Chrono-Nutrition and Cardiometabolic Health (2019) · cites this paper as [59]
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- Authors
- Louzada, P. R., Sebollela, A., Scaramello, M. E., & Ferreira, S. T.
- Affiliations
- Universidade Federal do Rio de Janeiro (BR)
- Venue
- Biophysical Journal
- Publisher
- Elsevier BV
- Published
- 2003-11
- Volume
- 85
- Issue
- 5
- Pages
- 3255-3261
- ISSN
- 0006-3495 (ISSN-L)
- PMC
- PMC1303601
- OpenAlex
- W2061545094
- Cited by
- 27
- References
- 39
- Citations by year
- 2026: 1 · 2021: 1 · 2019: 1 · 2018: 1 · 2016: 1 · 2014: 1 · 2013: 1
- FWCI
- 0.80
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biophysical Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Keywords
- glutathione reductase, protein unfolding, molten globule, guanidine hydrochloride, protein dimerization, folding intermediates, quaternary structure, thiol reactivity, protein folding mechanism, yeast enzymes
- MeSH terms
- Circular Dichroism, Enzyme Activation, Enzyme Stability, Glutathione Reductase, Hot Temperature, Protein Conformation, Saccharomyces cerevisiae, Spectrometry, Fluorescence, Protein Structure, Secondary, Protein Folding, Dimerization, Guanidine
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