Reference : Cofactor binding modulates the conformational stabilities and unfolding patterns of NAD(...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/15743
Cofactor binding modulates the conformational stabilities and unfolding patterns of NAD(+)-dependent DNA ligases from Escherichia coli and Thermus scotoductus
English
Georlette, D. [> > > >]
Blaise, Vinciane [Université de Liège - ULg > Département des sciences et gestion de l'environnement > Département des sciences et gestion de l'environnement >]
Dohmen, C. [> > > >]
Bouillenne, Fabrice mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Damien, B. [> > > >]
Depiereux, E. [> > > >]
Gerday, Charles mailto [Université de Liège - ULg > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences) >]
Uversky, V. N. [> > > >]
Feller, Georges mailto [Université de Liège - ULg > Département des sciences de la vie > Labo de biochimie >]
12-Dec-2003
Journal of Biological Chemistry
Amer Soc Biochemistry Molecular Biology Inc
278
50
49945-49953
Yes (verified by ORBi)
International
0021-9258
Bethesda
[en] DNA ligases are important enzymes required for cellular processes such as DNA replication, recombination, and repair. NAD(+)-dependent DNA ligases are essentially restricted to eubacteria, thus constituting an attractive target in the development of novel antibiotics. Although such a project might involve the systematic testing of a vast number of chemical compounds, it can essentially gain from the preliminary deciphering of the conformational stability and structural perturbations associated with the formation of the catalytically active adenylated enzyme. We have, therefore, investigated the adenylation-induced conformational changes in the mesophilic Escherichia coli and thermophilic Thermus scotoductus NAD(+)-DNA ligases, and the resistance of these enzymes to thermal and chemical (guanidine hydrochloride) denaturation. Our results clearly demonstrate that anchoring of the cofactor induces a conformational rearrangement within the active site of both mesophilic and thermophilic enzymes accompanied by their partial compaction. Furthermore, the adenylation of enzymes increases their resistance to thermal and chemical denaturation, establishing a thermodynamic link between cofactor binding and conformational stability enhancement. Finally, guanidine hydrochloride-induced unfolding of NAD(+)-dependent DNA ligases is shown to be a complex process that involves accumulation of at least two equilibrium intermediates, the molten globule and its precursor.
http://hdl.handle.net/2268/15743
10.1074/jbc.M307761200

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