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Thermodynamic basis for the stabilities of three CutA1s from Pyrococcus horikoshii,Thermus thermophilus, and Oryza sativa, with unusually high denaturation temperatures.
Biochemistry. 2008 Jan 15; 47(2):721-30.B

Abstract

In order to elucidate the stabilization mechanism of CutA1 from Pyrococcus horikoshii (PhCutA1) with a denaturation temperature of nearly 150 degrees C, GuHCl denaturation and heat denaturation were examined at neutral and acidic pHs. As a comparison, CutA1 proteins from Thermus thermophilus (TtCutA1) and Oryza sativa (OsCutA1) were also examined, which have lower optimum growth temperatures of 75 and 28 degrees C, respectively, than that (98 degrees C) of P. horikoshii. GuHCl-induced unfolding and refolding curves of the three proteins showed hysteresis effects due to an unusually slow unfolding rate. The midpoints of refolding for PhCutA1, TtCutA1 and OsCutA1 were 5.7 M, 3.3 M, and 2.3 M GuHCl, respectively, at pH 8.0 and 37 degrees C. DSC experiments with TtCutA1 and OsCutA1 showed that the denaturation temperatures were remarkably high, 112.8 and 97.3 degrees C, respectively, at pH 7.0 and that the good heat reversibility was amenable to thermodynamic analyses. At acidic pH, TtCutA1 showed higher stability to both heat and denaturant than PhCutA1. Combined with the data for DSC and denaturant denaturation, the unfolding Gibbs energy of PhCutA1 could be depicted as a function of temperature. It was experimentally revealed that (1) the unusually high stability of PhCutA1 basically originates from a common trimer structure of the three proteins, (2) the stability of PhCutA1 is superior to those of the other two CutA1s over all temperatures above 0 degrees C at neutral pH, due to the decrease in both enthalpy and entropy, and (3) ion pairs of PhCutA1 contribute to the unusually high stability at neutral pH.

Authors+Show Affiliations

RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18154307

Citation

Sawano, Masahide, et al. "Thermodynamic Basis for the Stabilities of Three CutA1s From Pyrococcus horikoshii,Thermus Thermophilus, and Oryza Sativa, With Unusually High Denaturation Temperatures." Biochemistry, vol. 47, no. 2, 2008, pp. 721-30.
Sawano M, Yamamoto H, Ogasahara K, et al. Thermodynamic basis for the stabilities of three CutA1s from Pyrococcus horikoshii,Thermus thermophilus, and Oryza sativa, with unusually high denaturation temperatures. Biochemistry. 2008;47(2):721-30.
Sawano, M., Yamamoto, H., Ogasahara, K., Kidokoro, S., Katoh, S., Ohnuma, T., Katoh, E., Yokoyama, S., & Yutani, K. (2008). Thermodynamic basis for the stabilities of three CutA1s from Pyrococcus horikoshii,Thermus thermophilus, and Oryza sativa, with unusually high denaturation temperatures. Biochemistry, 47(2), 721-30.
Sawano M, et al. Thermodynamic Basis for the Stabilities of Three CutA1s From Pyrococcus horikoshii,Thermus Thermophilus, and Oryza Sativa, With Unusually High Denaturation Temperatures. Biochemistry. 2008 Jan 15;47(2):721-30. PubMed PMID: 18154307.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Thermodynamic basis for the stabilities of three CutA1s from Pyrococcus horikoshii,Thermus thermophilus, and Oryza sativa, with unusually high denaturation temperatures. AU - Sawano,Masahide, AU - Yamamoto,Hitoshi, AU - Ogasahara,Kyoko, AU - Kidokoro,Shun-ichi, AU - Katoh,Shizue, AU - Ohnuma,Takayuki, AU - Katoh,Etsuko, AU - Yokoyama,Shigeyuki, AU - Yutani,Katsuhide, Y1 - 2007/12/22/ PY - 2007/12/25/pubmed PY - 2008/3/4/medline PY - 2007/12/25/entrez SP - 721 EP - 30 JF - Biochemistry JO - Biochemistry VL - 47 IS - 2 N2 - In order to elucidate the stabilization mechanism of CutA1 from Pyrococcus horikoshii (PhCutA1) with a denaturation temperature of nearly 150 degrees C, GuHCl denaturation and heat denaturation were examined at neutral and acidic pHs. As a comparison, CutA1 proteins from Thermus thermophilus (TtCutA1) and Oryza sativa (OsCutA1) were also examined, which have lower optimum growth temperatures of 75 and 28 degrees C, respectively, than that (98 degrees C) of P. horikoshii. GuHCl-induced unfolding and refolding curves of the three proteins showed hysteresis effects due to an unusually slow unfolding rate. The midpoints of refolding for PhCutA1, TtCutA1 and OsCutA1 were 5.7 M, 3.3 M, and 2.3 M GuHCl, respectively, at pH 8.0 and 37 degrees C. DSC experiments with TtCutA1 and OsCutA1 showed that the denaturation temperatures were remarkably high, 112.8 and 97.3 degrees C, respectively, at pH 7.0 and that the good heat reversibility was amenable to thermodynamic analyses. At acidic pH, TtCutA1 showed higher stability to both heat and denaturant than PhCutA1. Combined with the data for DSC and denaturant denaturation, the unfolding Gibbs energy of PhCutA1 could be depicted as a function of temperature. It was experimentally revealed that (1) the unusually high stability of PhCutA1 basically originates from a common trimer structure of the three proteins, (2) the stability of PhCutA1 is superior to those of the other two CutA1s over all temperatures above 0 degrees C at neutral pH, due to the decrease in both enthalpy and entropy, and (3) ion pairs of PhCutA1 contribute to the unusually high stability at neutral pH. SN - 0006-2960 UR - https://www.unboundmedicine.com/medline/citation/18154307/Thermodynamic_basis_for_the_stabilities_of_three_CutA1s_from_Pyrococcus_horikoshiiThermus_thermophilus_and_Oryza_sativa_with_unusually_high_denaturation_temperatures_ L2 - https://doi.org/10.1021/bi701761m DB - PRIME DP - Unbound Medicine ER -