DOI: 10.1042/bj2990347. The significance of denaturant titrations of protein stability: a comparison of rat and baker's yeast cytochrome <i>c</i> and their site-directed asparagine-52-to-isoleucine mutants

The significance of denaturant titrations of protein stability: a comparison of rat and baker's yeast cytochrome c and their site-directed asparagine-52-to-isoleucine mutants

10.1042/bj2990347

Crossref journal-article

Portland Press Ltd.

Biochemical Journal (288)

Abstract

The residue asparagine-52 of rat cytochrome c and baker's yeast iso-1-cytochrome c was mutated to isoleucine by site-directed mutagenesis, and the unfolding of the wild-type and mutant proteins in urea or guanidinium chloride solutions was studied. Whereas the yeast mutant cytochrome unfolded in 4-7 M urea with a rate constant (k) of 1.7 x 10(-2) s-1, the rat mutant protein unfolded with k = 5.0 x 10(-2) s-1, followed by a slow partial refolding with k = 5.0 x 10(-4) s-1. Denaturant titrations indicated that the mutation increased the stability of the yeast cytochrome by 6.3 kJ (1.5 kcal)/mol, while it decreased that of the rat protein by 11.7 kJ (2.8 kcal)/mol. These results probably reflect structural differences between yeast iso-1 and vertebrate cytochromes c in the vicinity of the Asn-52 side chain.

Bibliography

Koshy, T. I., Luntz, T. L., Plotkin, B., Schejter, A., & Margoliash, E. (1994). The significance of denaturant titrations of protein stability: a comparison of rat and bakerâs yeast cytochrome c and their site-directed asparagine-52-to-isoleucine mutants. Biochemical Journal, 299(2), 347â350.

Authors 5

T I Koshy (first)
T L Luntz (additional)
B Plotkin (additional)
A Schejter (additional)
E Margoliash (additional)

References 0 Referenced 10

None

Dates

Type	When
Created	10 years ago (Aug. 10, 2015, 5:37 p.m.)
Deposited	3 years, 9 months ago (Nov. 23, 2021, 5:37 p.m.)
Indexed	1 year, 9 months ago (Oct. 31, 2023, 6:20 a.m.)
Issued	31 years, 4 months ago (April 15, 1994)
Published	31 years, 4 months ago (April 15, 1994)
Published Print	31 years, 4 months ago (April 15, 1994)

Funders 0

None

BibTeX

@article{Koshy_1994, title={The significance of denaturant titrations of protein stability: a comparison of rat and baker’s yeast cytochrome c and their site-directed asparagine-52-to-isoleucine mutants}, volume={299}, ISSN={1470-8728}, url={http://dx.doi.org/10.1042/bj2990347}, DOI={10.1042/bj2990347}, number={2}, journal={Biochemical Journal}, publisher={Portland Press Ltd.}, author={Koshy, T I and Luntz, T L and Plotkin, B and Schejter, A and Margoliash, E}, year={1994}, month=apr, pages={347–350} }

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