DOI: 10.1002/pro.5560031110. Protein denaturation with guanidine hydrochloride or urea provides a different estimate of stability depending on the contributions of electrostatic interactions

Protein denaturation with guanidine hydrochloride or urea provides a different estimate of stability depending on the contributions of electrostatic interactions

10.1002/pro.5560031110

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractThe objective of this study was to address the question of whether or not urea and guanidine hydrochloride (GdnHCl) give the same estimates of the stability of a particular protein. We previously suspected that the estimates of protein stability from GdnHCl and urea denaturation data might differ depending on the electrostatic interactions stabilizing the proteins. Therefore, 4 coiled‐coil analogs were designed, where the number of intrachain and interchain electrostatic attractions (A) were systematically changed to repulsions (R): 20A, 15A5R, 10A10R, and 20R. The GdnHCl denaturation data showed that the 4 coiled‐coil analogs, which had electrostatic interactions ranging from 20 attractions to 20 repulsions, had very similar [GdnHCl]1/2 values (average of ⋍3.5 M) and, as well, their ΔΔGu values were very close to 0 (0.2 kcal/mol). In contrast, urea denaturation showed that the [urea]1/2 values proportionately decreased with the stepwise change from 20 electrostatic attractions to 20 repulsions (20A, 7.4 M; 15A5R, 5.4 M; 10A10R, 3.2 M; and 20R, 1.4 M), and the ΔΔGu values correspondingly increased with the increasing differences in electrostatic interactions (20A – 15A5R, 1.5 kcal/mol; 20A – 10A10R, 3.7 kcal/mol; and 20A – 20R, 5.8 kcal/mol). These results indicate that the ionic nature of GdnHCl masks electrostatic interactions in these model proteins, a phenomenon that was absent when the uncharged urea was used. Thus, GdnHCl and urea denaturations may give vastly different estimates of protein stability, depending on how important electrostatic interactions are to the protein.

Bibliography

Monera, O. D., Kay, C. M., & Hodges, R. S. (1994). Protein denaturation with guanidine hydrochloride or urea provides a different estimate of stability depending on the contributions of electrostatic interactions. Protein Science, 3(11), 1984â1991. Portico.

Authors 3

Oscar D. Monera (first)
Cyril M. Kay (additional)
Robert S. Hodges (additional)

References 33 Referenced 302

10.1016/S0021-9258(18)33605-6 / J Biol Chem / Estimation of the free energy of stabilization of ribonuclease A, lysozyme, α‐lactalbumin, and myoglobin by Ahmad F (1982)
10.1021/bi00421a015
10.1021/bi00444a001
{'key': 'e_1_2_1_5_1', 'volume-title': 'Proteins', 'author': 'Creighton T.', 'year': '1993'} / Proteins by Creighton T. (1993)
10.1021/bi00483a001
10.1021/bi00227a002
10.1021/bi00140a005
10.1016/S0021-9258(20)79739-5
10.1016/S0021-9258(18)90686-1
10.1021/bi00699a005
10.1016/0022-2836(92)90963-K
10.1073/pnas.84.19.6663
10.1126/science.8235609
10.1021/bi00082a021
10.1021/bi00179a010
10.1016/S0021-9258(19)36502-0 / J Biol Chem / Comparison of anti‐parallel and parallel two‐stranded α‐helical coiled‐coils: Design, synthesis, and characterization by Monera OD (1993)
10.3109/10409237509102551
10.1016/0076-6879(86)31045-0
10.1016/0968-0004(90)90124-T
10.1021/bi00462a019
10.1016/0003-9861(81)90209-5
10.1016/S0065-3233(08)60460-X
10.1021/ja01089a028
10.1021/ja00836a027
10.1016/0022-2836(91)90117-O
10.1021/bi00421a014
10.1016/S0021-9258(18)83544-X / J Biol Chem / Probing the determinants of protein folding and stability with amino acid substitutions by Shortle D. (1989)
10.1016/S0065-3233(08)60241-7
10.1021/bi00072a001
10.1016/S0021-9258(18)45932-7 / J Biol Chem / Synthetic model proteins: Positional effects of interchain hydrophobic interactions on stability of two‐stranded α‐helical coiled‐coils by Zhou NE (1992)
10.1021/bi00140a008
10.1006/jmbi.1994.1250
10.1111/j.1399-3011.1992.tb00290.x

Dates

Type	When
Created	16 years, 6 months ago (Feb. 9, 2009, 8:27 p.m.)
Deposited	1 year, 10 months ago (Oct. 24, 2023, 4:12 p.m.)
Indexed	5 days, 15 hours ago (Aug. 29, 2025, 6:06 a.m.)
Issued	30 years, 10 months ago (Nov. 1, 1994)
Published	30 years, 10 months ago (Nov. 1, 1994)
Published Online	16 years, 8 months ago (Dec. 31, 2008)
Published Print	30 years, 10 months ago (Nov. 1, 1994)

Funders 0

None

BibTeX

@article{Monera_1994, title={Protein denaturation with guanidine hydrochloride or urea provides a different estimate of stability depending on the contributions of electrostatic interactions}, volume={3}, ISSN={1469-896X}, url={http://dx.doi.org/10.1002/pro.5560031110}, DOI={10.1002/pro.5560031110}, number={11}, journal={Protein Science}, publisher={Wiley}, author={Monera, Oscar D. and Kay, Cyril M. and Hodges, Robert S.}, year={1994}, month=nov, pages={1984–1991} }

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