Conformational Polymorphism of High MrGlutenin Subunits Detected by Transverse Urea Gradient Gel Electrophoresis

Lafiandra, D.; Turchetta, T.; D’Ovidio, R.; Anderson, Olin D.; Facchiano, Angelo; Colonna, Giovanni

doi:10.1006/jcrs.1999.0260

Cited by 15 publications

(10 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Multidomain proteins may exhibit complex equilibrium transitions as different domains can unfold independently. Results from studies carried out on small globular proteins demonstrated that the data obtained from electrophoretic separations are consistent with those obtained with conventional and more sophisticated biophysical methods [70]. In an analogous manner, unfolding and refolding can also be studied by generating a temperature gradient across the gel [71][72][73].…”

Section: Gel-slab Electrophoresis In Transverse Denaturant Gradientsmentioning

confidence: 54%

Folding/unfolding/refolding of proteins: Present methodologies in comparison with capillary zone electrophoresis

Righetti

Verzola

2001

Electrophoresis

View full text Add to dashboard Cite

A series of techniques for monitoring protein folding/unfolding/misfolding equilibria are here assessed and compared with capillary zone electrophoresis (CZE). They include spectroscopic techniques, such as circular dichroism, intrinsic fluorescence, nuclear magnetic resonance, Fourier transform infrared and Raman spectroscopy, small-angle X-ray scattering, as well as techniques based on biological assays, such as limited proteolysis and immunochemical analysis of different conformational states. Some unusual probes, such as mass spectrometry for probing unfolding transitions, are also discussed. Size-exclusion chromatography is also evaluated in view of the fact that this technique, like all electrophoretic techniques, and unlike spectroscopic probes, which can only see an average signal in mixed populations, can indeed physically separate folded vs. unfolded macromolecules, especially in the case of slow equilibria. Particular emphasis is devoted to electrophoretic techniques, such as gel-slab electrophoresis in transverse urea or thermal gradients, and CZE. In the latter case, a number of applications are shown, demonstrating the excellent correlation of CZE with more traditional probes, such as intrinsic fluorescence monitoring. It is additionally shown that CZE can be used for measuring the deltaG degrees of unfolding over the pH scale, in good agreement with theoretical calculations on the electrostatic free energy of folding vs. pH, as calculated with a linearized Poisson-Boltzmann equation. Finally, it is demonstrated that CZE can probe also aggregate formation in the presence of helix-inducing agents, such as trifluorethanol.

show abstract

Section: Gel-slab Electrophoresis In Transverse Denaturant Gradientsmentioning

confidence: 54%

Folding/unfolding/refolding of proteins: Present methodologies in comparison with capillary zone electrophoresis

Righetti

Verzola

2001

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Secondly, we have shown that subunit 1Dy10 shows significantly higher stability than 1Dy12 when unfolded on transverse urea gradient gels (with a free energy of 7.4 compared with 5.1 Kcal/mol). This may indicate that deformation on mixing would require a greater energy input, although it is difficult to make direct extrapolations between the results obtained by electrophoresis in the presence of a detergent with the behaviour in dough [33]. These comparisons nevertheless suggest that individual features of subunits 1DÂ5 and 1Dy10 may independently contribute to dough strength.…”

Section: Good and Poor Quality Subunitsmentioning

confidence: 83%

Wheat glutenin subunits and dough elasticity: findings of the EUROWHEAT project

Shewry

Popineau²,

Lafiandra³

et al. 2000

Trends in Food Science & Technology

231

137

View full text Add to dashboard Cite

Detailed studies of wheat glutenin subunits have provided novel details of their molecular structures and interactions which allow the development of a model to explain their role in determining the visco-elastic properties of gluten and dough. The construction and analysis of near-isogenic and transgenic lines expressing novel subunit combinations or increased amounts of specific subunits allows differences in gluten properties to be related to the structures and properties of individual subunits, with potential benefits for the production of cultivars with improved properties for food processing or novel end users #

show abstract

“…In addition to containing less cysteine residues, subunit 1Bx20 also appears to differ from all other HMW subunits in its conformational stability, showing an unusual concave pattern when unfolded on transverse urea gels in the presence of SDS (Lafiandra et al 1999). It is possible, therefore, that the unusually low cysteine content and/or the low conformational stability could contribute to the association of subunit 1Bx20 with poor quality for pasta making.…”

Section: Introductionmentioning

confidence: 90%

Sequence and properties of HMW subunit 1Bx20 from pasta wheat (Triticum durum) which is associated with poor end use properties

et al. 2003

View full text Add to dashboard Cite

The gene encoding high-molecular-weight (HMW) subunit 1Bx20 was isolated from durum wheat cv. Lira. It encodes a mature protein of 774 amino acid residues with an M(r) of 83,913. Comparison with the sequence of subunit 1Bx7 showed over 96% identity, the main difference being the substitution of two cysteine residues in the N-terminal domain of subunit 1Bx7 with tyrosine residues in 1Bx20. Comparison of the structures and stabilities of the two subunits purified from wheat using Fourier-transform infra-red and circular dichroism spectroscopy showed no significant differences. However, incorporation of subunit 1Bx7 into a base flour gave increased dough strength and stability measured by Mixograph analysis, while incorporation of subunit 1Bx20 resulted in small positive or negative effects on the parameters measured. It is concluded that the different effects of the two subunits could relate to the differences in their cysteine contents, thereby affecting the cross-linking and hence properties of the glutenin polymers.

show abstract

Conformational Polymorphism of High MrGlutenin Subunits Detected by Transverse Urea Gradient Gel Electrophoresis

Cited by 15 publications

References 16 publications

Folding/unfolding/refolding of proteins: Present methodologies in comparison with capillary zone electrophoresis

Folding/unfolding/refolding of proteins: Present methodologies in comparison with capillary zone electrophoresis

Wheat glutenin subunits and dough elasticity: findings of the EUROWHEAT project

Sequence and properties of HMW subunit 1Bx20 from pasta wheat (Triticum durum) which is associated with poor end use properties

Contact Info

Product

Resources

About