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Thermal stability and atomic-resolution crystal structure of the Bacillus caldolyticus cold shock protein.
J Mol Biol. 2000 Apr 07; 297(4):975-88.JM

Abstract

The bacterial cold shock proteins are small compact beta-barrel proteins without disulfide bonds, cis-proline residues or tightly bound cofactors. Bc-Csp, the cold shock protein from the thermophile Bacillus caldolyticus shows a twofold increase in the free energy of stabilization relative to its homolog Bs-CspB from the mesophile Bacillus subtilis, although the two proteins differ by only 12 out of 67 amino acid residues. This pair of cold shock proteins thus represents a good system to study the atomic determinants of protein thermostability. Bs-CspB and Bc-Csp both unfold reversibly in cooperative transitions with T(M) values of 49.0 degrees C and 77.3 degrees C, respectively, at pH 7.0. Addition of 0.5 M salt stabilizes Bs-CspB but destabilizes Bc-Csp. To understand these differences at the structural level, the crystal structure of Bc-Csp was determined at 1.17 A resolution and refined to R=12.5% (R(free)=17.9%). The molecular structures of Bc-Csp and Bs-CspB are virtually identical in the central beta-sheet and in the binding region for nucleic acids. Significant differences are found in the distribution of surface charges including a sodium ion binding site present in Bc-Csp, which was not observed in the crystal structure of the Bs-CspB. Electrostatic interactions are overall favorable for Bc-Csp, but unfavorable for Bs-CspB. They provide the major source for the increased thermostability of Bc-Csp. This can be explained based on the atomic-resolution crystal structure of Bc-Csp. It identifies a number of potentially stabilizing ionic interactions including a cation-binding site and reveals significant changes in the electrostatic surface potential.

Authors+Show Affiliations

Forschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Robert-Roessle-Str. 10, Berlin, D-13125, Germany.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

10736231

Citation

Mueller, U, et al. "Thermal Stability and Atomic-resolution Crystal Structure of the Bacillus Caldolyticus Cold Shock Protein." Journal of Molecular Biology, vol. 297, no. 4, 2000, pp. 975-88.
Mueller U, Perl D, Schmid FX, et al. Thermal stability and atomic-resolution crystal structure of the Bacillus caldolyticus cold shock protein. J Mol Biol. 2000;297(4):975-88.
Mueller, U., Perl, D., Schmid, F. X., & Heinemann, U. (2000). Thermal stability and atomic-resolution crystal structure of the Bacillus caldolyticus cold shock protein. Journal of Molecular Biology, 297(4), 975-88.
Mueller U, et al. Thermal Stability and Atomic-resolution Crystal Structure of the Bacillus Caldolyticus Cold Shock Protein. J Mol Biol. 2000 Apr 7;297(4):975-88. PubMed PMID: 10736231.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Thermal stability and atomic-resolution crystal structure of the Bacillus caldolyticus cold shock protein. AU - Mueller,U, AU - Perl,D, AU - Schmid,F X, AU - Heinemann,U, PY - 2000/3/29/pubmed PY - 2000/5/8/medline PY - 2000/3/29/entrez SP - 975 EP - 88 JF - Journal of molecular biology JO - J Mol Biol VL - 297 IS - 4 N2 - The bacterial cold shock proteins are small compact beta-barrel proteins without disulfide bonds, cis-proline residues or tightly bound cofactors. Bc-Csp, the cold shock protein from the thermophile Bacillus caldolyticus shows a twofold increase in the free energy of stabilization relative to its homolog Bs-CspB from the mesophile Bacillus subtilis, although the two proteins differ by only 12 out of 67 amino acid residues. This pair of cold shock proteins thus represents a good system to study the atomic determinants of protein thermostability. Bs-CspB and Bc-Csp both unfold reversibly in cooperative transitions with T(M) values of 49.0 degrees C and 77.3 degrees C, respectively, at pH 7.0. Addition of 0.5 M salt stabilizes Bs-CspB but destabilizes Bc-Csp. To understand these differences at the structural level, the crystal structure of Bc-Csp was determined at 1.17 A resolution and refined to R=12.5% (R(free)=17.9%). The molecular structures of Bc-Csp and Bs-CspB are virtually identical in the central beta-sheet and in the binding region for nucleic acids. Significant differences are found in the distribution of surface charges including a sodium ion binding site present in Bc-Csp, which was not observed in the crystal structure of the Bs-CspB. Electrostatic interactions are overall favorable for Bc-Csp, but unfavorable for Bs-CspB. They provide the major source for the increased thermostability of Bc-Csp. This can be explained based on the atomic-resolution crystal structure of Bc-Csp. It identifies a number of potentially stabilizing ionic interactions including a cation-binding site and reveals significant changes in the electrostatic surface potential. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/10736231/Thermal_stability_and_atomic_resolution_crystal_structure_of_the_Bacillus_caldolyticus_cold_shock_protein_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(00)93602-7 DB - PRIME DP - Unbound Medicine ER -