title: Temperature modulates binding specificity and affinity of the d-trehalose/d-maltose-binding protein from the hyperthermophilic archaeon Thermococcus litoralis.
abstract: We investigated the effect of temperature on the binding specificity of the recombinant d-trehalose/d-maltose-binding protein from the hyperthermophilic archaeon Thermococcus litoralis (TMBP). Importantly, we found that TMBP can bind d-glucose (Glc). The Glc binding was characterized by means of fluorescence spectroscopy in the temperature range of 25 degrees C-85 degrees C. Our results show that at 25 degrees C the binding of Glc to TMBP is well represented by a bimodal model with apparent K(d) of 20 muM and approximately 3-8 mM for the first and the second binding step, respectively. At 60 degrees C the binding of Glc to TMBP is represented by a simple hyperbolic model with an apparent K(d) value of about 40 muM. Finally, at 85 degrees C Glc did not bind to TMBP. Molecular dynamics (MD) simulations were used to shed light on the molecular mechanism of the Glc binding. Our results suggest that after proper fluorescent labeling TMBP can be used as a highly thermostable and non-consuming analyte biosensor for monitoring the level of glucose in fluids (e.g. human blood) where other sugars are not present.
authors: Faculty of Mathematics and Physics, Institute of Physics, Charles University, Ke Karlovu 5, 121 16 Prague, Czech Republic. herman@karlov.mff.cuni.cz
authors: Petr
authors: P
authors: Herman
authors: Ivan
authors: I
authors: Barvik
authors: Maria
authors: M
authors: Staiano
authors: Annalisa
authors: A
authors: Vitale
authors: Jaroslav
authors: J
authors: Vecer
authors: Mose'
authors: M
authors: Rossi
authors: Sabato
authors: S
authors: D'Auria
mesh_terms: D019843:Archaeal Proteins / Q000378:metabolism*; D008320:Maltose / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013696:Temperature; D019712:Thermococcus / Q000302:isolation & purification / Q000378:metabolism*; D014199:Trehalose / Q000378:metabolism*
chemical_list: D019843:Archaeal Proteins; D008320:Maltose; D014199:Trehalose