Circular Dichroism Evidence for the Presence of Burst-Phase Intermediates on the Conformational Folding Pathway of Ribonuclease A†

Biochemistry ◽  
1996 ◽  
Vol 35 (31) ◽  
pp. 10125-10133 ◽  
Author(s):  
Walid A. Houry ◽  
David M. Rothwarf ◽  
Harold A. Scheraga
Keyword(s):  
Circular Dichroism ◽  
Ribonuclease A ◽  
Folding Pathway ◽  
Burst Phase ◽  
Circular Dichroïsm

Folding Pathway of Escherichia coli Ribonuclease HI: A Circular Dichroism, Fluorescence, and NMR Study

Biochemistry ◽  
1995 ◽  
Vol 34 (51) ◽  
pp. 16552-16562 ◽  
Author(s):  
Kazuhiko Yamasaki ◽  
Kyoko Ogasahara ◽  
Katsuhide Yutani ◽  
Motohisa Oobatake ◽  
Shigenori Kanaya
Keyword(s):  
Escherichia Coli ◽  
Circular Dichroism ◽  
Folding Pathway ◽  
Nmr Study ◽  
Circular Dichroïsm

scholarly journals Bis-Lactam Peptide [i, i+4]-Stapling with α-Methylated Thialysines

Molecules ◽  
2020 ◽  
Vol 25 (19) ◽  
pp. 4506
Author(s):  
Bo Wu ◽  
Weiping Zheng
Keyword(s):  
Circular Dichroism ◽  
Ribonuclease A ◽  
Rnase A ◽  
Peptide Sequence ◽  
Stability Assessment ◽  
Model Peptide ◽  
Stapled Peptides ◽  
Stapled Peptide ◽  
Proteolytic Stability ◽  
Circular Dichroïsm

Four bis-lactam [i, i+4]-stapled peptides with d- or l-α-methyl-thialysines were constructed on a model peptide sequence derived from p110α[E545K] and subjected to circular dichroism (CD) and proteolytic stability assessment, alongside the corresponding bis-lactam [i, i+4]-stapled peptide with l-thialysine. The % α-helicity values of these four stapled peptides were found to be largely comparable to each other yet greater than that of the stapled peptide with l-thialysine. An l-α-methyl-thialysine-stapled peptide built on a model peptide sequence derived from ribonuclease A (RNase A) was also found to exhibit a greater % α-helicity than its l-thialysine-stapled counterpart. Moreover, a greater proteolytic stability was demonstrated for the l-α-methyl-thialysine-stapled p110α[E545K] and RNase A peptides than that of their respective l-thialysine-stapled counterparts.

scholarly journals Folding mechanism of canine milk lysozyme studied by circular dichroism and fluorescence spectroscopy

2003 ◽  
Vol 17 (2-3) ◽  
pp. 183-193 ◽  
Author(s):  
Masaharu Nakao ◽  
Munehito Arai ◽  
Takumi Koshiba ◽  
Katsutoshi Nitta ◽  
Kunihiro Kuwajima
Keyword(s):  
Circular Dichroism ◽  
Fluorescence Spectroscopy ◽  
Molten Globule ◽  
Guanidine Hydrochloride ◽  
Folding Intermediates ◽  
Equilibrium Unfolding ◽  
Burst Phase ◽  
The Relationship ◽  
Kinetics Of ◽  
Circular Dichroïsm

We have studied the guanidine hydrochloride‒induced equilibrium unfolding and the kinetics of refolding of canine milk lysozyme by circular dichroism and fluorescence spectroscopy. The thermodynamic analysis of the equilibrium unfolding measured by circular dichroism and fluorescence has shown that unfolding is represented by a three‒state mechanism and that the intermediate state of canine milk lysozyme is remarkably more stable than the intermediates observed in other lysozyme and α-lactalbumin. In the kinetic refolding of this protein, there are at least two kinetic intermediates; a burst=phase intermediate accumulated within the dead time (4 ms) of the measurement and an intermediate that has been observed during the kinetics with a rate constant of 10–20 s–1after the burst phase. This result is apparently in contrast with those previously observed in the kinetic refolding of α‒lactalbumin and equine lysozyme that show only the burst‒phase intermediate. The relationship between the extraordinarily stable equilibrium molten globule and the kinetic folding intermediates will be discussed.

The quantitative comparison of the effects of denaturants on the stability of proteins

1986 ◽  
Vol 64 (10) ◽  
pp. 993-998 ◽  
Author(s):  
James A. Thomson ◽  
Charles C. Bigelow
Keyword(s):  
Circular Dichroism ◽  
Specific Binding ◽  
Ribonuclease A ◽  
Quantitative Comparison ◽  
Difference Spectroscopy ◽  
Guanidinium Chloride ◽  
Binding Capability ◽  
The Stability ◽  
Circular Dichroïsm

A novel quantitative comparison of denaturants involving the complete reversible unfolding of proteins is presented. Ribonuclease A was denatured with guanidinium chloride in the presence of low fixed concentrations of various partial denaturants, with the unfolding process being monitored by circular dichroism and difference spectroscopy. The major advantage of this method is that it allows a direct quantitative comparison of the effects of denaturants on the stability of proteins. The effect on the stability of ribonuclease A was shown to be linearly dependent upon the concentration of denaturant. An investigation of the constitutive ions of salts revealed that their effects were additive only in the case of salts that have no specific binding capability. This method can also be useful in detecting the specific binding of salts.

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