PMID: 12787677Jun 6, 2003Paper

Effects of the difference in the unfolded-state ensemble on the folding of Escherichia coli dihydrofolate reductase

Journal of Molecular Biology
Munehito AraiM Iwakura

Abstract

The unfolded state of a protein is an ensemble of a large number of conformations ranging from fully extended to compact structures. To investigate the effects of the difference in the unfolded-state ensemble on protein folding, we have studied the structure, stability, and folding of "circular" dihydrofolate reductase (DHFR) from Escherichia coli in which the N and C-terminal regions are cross-linked by a disulfide bond, and compared the results with those of disulfide-reduced "linear" DHFR. Equilibrium studies by circular dichroism, difference absorption spectra, solution X-ray scattering, and size-exclusion chromatography show that whereas the native structures of both proteins are essentially the same, the unfolded state of circular DHFR adopts more compact conformations than the unfolded state of the linear form, even with the absence of secondary structure. Circular DHFR is more stable than linear DHFR, which may be due to the decrease in the conformational entropy of the unfolded state as a result of circularization. Kinetic refolding measurements by stopped-flow circular dichroism and fluorescence show that under the native conditions both proteins accumulate a burst-phase intermediate having the same structures and bot...Continue Reading

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Citations

Jun 26, 2007·Bioscience, Biotechnology, and Biochemistry·Hayuki SugimotoAkiyoshi Tanaka
Aug 18, 2004·Proceedings of the National Academy of Sciences of the United States of America·Jonathan E KohnKevin W Plaxco
May 13, 2015·Proceedings of the National Academy of Sciences of the United States of America·Herschel M WatkinsKevin W Plaxco
Aug 30, 2005·Critical Reviews in Biochemistry and Molecular Biology·Evan R McCarneyKevin W Plaxco
Feb 1, 2007·The Journal of Biological Chemistry·Hisashi TakahashiMasahiro Iwakura
Mar 3, 2006·The Journal of Biological Chemistry·Masahiro IwakuraKunihiko Gekko
Jan 4, 2018·Bioorganic & Medicinal Chemistry·Rui-Yang ZhangJon-Paul Bingham
Jun 9, 2021·The Journal of Physical Chemistry. B·Beatriz Fernández Del Río, Antonio Rey
Oct 13, 2004·Biochemistry·Wei Yuan Yang, Martin Gruebele
Nov 23, 2005·Biochemistry·Irina BezsonovaJulie D Forman-Kay

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