Molecular adaptation strategies to high temperature and thermal denaturation mechanism of the D-trehalose/D-maltose-binding protein from the hyperthermophilic archaeon Thermococcus litoralis.
The D-trehalose/D-maltose-binding protein (TMBP), a monomeric protein of 48 kDa, is one component of the trehalose and maltose uptake system. In the hyperthermophilic archaeon T. litoralis this is mediated by a protein-dependent ATP-binding cassette system transporter. The gene coding for a thermostable TMBP from the archaeon T. litoralis has been cloned, and the recombinant protein has been expressed in E. coli. The recombinant TMBP has been purified to homogeneity and characterized. It exhibits the same functional and structural properties as the native one. In fact, it is highly thermostable and binds both trehalose and maltose with high affinity. In this work we used differential scanning calorimetry studies together with a detailed analysis, at the molecular level, of the three-dimensional protein structure to shed light on the basis of the high thermostability exhibited by the recombinant TMBP from the archaeon T. litoralis. The obtained data suggest that the presence of trehalose does not change the overall mechanism of the denaturation of this protein but it selectively modifies the stability of the TMBP structural domains.
Fessas
Dimitrios
D
Department of Food Science, Technology and Microbiology, University of Milan, Milan, Italy.
Staiano
Maria
M
Barbiroli
Alberto
A
Marabotti
Anna
A
Schiraldi
Alberto
A
Varriale
Antonio
A
Rossi
Mose'
M
D'Auria
Sabato
S
D000220:Adaptation, Biological; D002152:Calorimetry, Differential Scanning; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018360:Crystallography, X-Ray; D058619:Maltose-Binding Proteins; D008958:Models, Molecular; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D013696:Temperature; D019712:Thermococcus / Q000201:enzymology*; D014199:Trehalose / Q000737:chemistry* / Q000378:metabolism*
D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D014199:Trehalose