Thermostability of Multidomain Proteins: Properties of Chimaeric Elongation Factors EF-Tu Composed of Domains of Mesophilic-Escherichia coli EF-Tu and Thermophilic-Bacillus stearothermophilus EF-Tu

0191634 - UMG-J 20033142 RIV CZ eng J - Journal Article
Šanderová, Hana - Hůlková, Marta - Maloň, Petr - Jonák, Jiří
Thermostability of Multidomain Proteins: Properties of Chimaeric Elongation Factors EF-Tu Composed of Domains of Mesophilic-Escherichia coli EF-Tu and Thermophilic-Bacillus stearothermophilus EF-Tu.
Chemické listy. Roč. 97, č. 5 (2003), s. 308-309. ISSN 0009-2770. E-ISSN 1213-7103
R&D Projects: GA ČR GA204/98/0863; GA ČR GA303/02/0689
Institutional research plan: CEZ:AV0Z4055905; CEZ:AV0Z5052915
Keywords : elongation factor EF-Tu, thermostability, chimeric protein
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 0.345, year: 2003

The EF-Tu proteins from E. coli and B. stearothermophilus were examined by the chimaerization approach to evaluate the contribution of the domains to the thermostability of these proteins. Molecules of EF-Tus were genetically dissected into three corresponding domains and the domains combined to form chimaeric EF-Tu proteins. The resulting six recombinant mesophile/thermophile chimaeric EF-Tus, together with the recombinant E. coli and B. stearothermophilus EF-Tus and isolated G-domains, were characterized with regard to GDP and GTP binding activity, intrinsic GTPase activity and thermostability. The thermostability was measured both as the maintenance, at increasing temperatures, of a defined functional state by the ability to bind GDP and GTP, and to hydrolyze GTP and, independently, using CD spectroscopy, as the maintenance of the alfa-helix content.
Permanent Link: http://hdl.handle.net/11104/0087372