Reference : Structural Determinants of Specificity and Catalytic Mechanism in mammalian 25-kDa Th...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/149136
Structural Determinants of Specificity and Catalytic Mechanism in mammalian 25-kDa Thiamine Triphosphatase
English
Delvaux, David [Université de Liège - ULg > GIGA-Neurosciences > > >]
Kerff, Frédéric mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Murty, Mamidanna R.V.S. []
Lakaye, Bernard mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique >]
Czerniecki, Jan []
Kohn, Grégory mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique >]
Wins, Pierre []
Herman, Raphaël mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Gabelica, Valérie mailto [Université de Liège - ULg > Département de chimie (sciences) > GIGA-R : Laboratoire de spectrométrie de masse (L.S.M.) >]
Heuze, Fabien []
Tordoir, Xavier []
Marée, Raphaël mailto [Université de Liège - ULg > > GIGA-Management : Plateforme bioinformatique >]
Matagne, André mailto [Université de Liège - ULg > Département des sciences de la vie > Enzymologie et repliement des protéines >]
Charlier, Paulette mailto [Université de Liège - ULg > Département des sciences de la vie > Cristallographie des macromolécules biologiques >]
De Pauw, Edwin mailto [Université de Liège - ULg > Département de chimie (sciences) > GIGA-R : Laboratoire de spectrométrie de masse (L.S.M.) >]
Bettendorff, Lucien mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique >]
2013
Biochimica et Biophysica Acta - General Subjects
Elsevier Science
1830
4513-4523
Yes (verified by ORBi)
International
0304-4165
1872-8006
Amsterdam
The Netherlands
[en] thiamine ; triphosphate tunnel metalloenzymes ; CYTH ; tripolyphosphate ; divalent cation
[en] Background: Thiamine triphosphate (ThTP) is present in most organisms and might be involved in intracellular signaling. In mammalian cells, the cytosolic ThTP level is controlled by a specific thiamine triphosphatase (ThTPase), belonging to the CYTH superfamily of proteins. CYTH proteins are present in all superkingdoms of life and act on various triphosphorylated substrates. Methods: Using crystallography, mass spectrometry and mutational analysis, we identified the key structural determinants of the high specificity and catalytic efficiency of mammalian ThTPase. Results: Triphosphate binding requires three conserved arginines while the catalytic mechanism relies on an unusual lysine-tyrosine dyad. By docking of the ThTP molecule in the active site, we found that Trp-53 should interact with the thiazole part of the substrate molecule, thus playing a key role in substrate recognition and specificity. Sea anemone and zebrafish CYTH proteins, which retain the corresponding Trp residue, are also specific ThTPases. Surprisingly, the whole chromosome region containing the ThTPase gene is lost in birds. Conclusion: The specificity for ThTP is linked to a stacking interaction between the thiazole heterocycle of thiamine and a tryptophan residue. The latter likely plays a key role in the secondary acquisition of ThTPase activity in early metazoan CYTH enzymes, in the lineage leading from cnidarians to mammals. General significance: We show that ThTPase activity is not restricted to mammals as previously thought but is an acquisition of early metazoans. This, and the identification of critically important residues, allows us to draw an evolutionary perspective of the CYTH family of proteins.
Giga-Neurosciences ; Centre d'Ingénierie des Protéines - CIP ; Giga-Systems Biology and Chemical Biology ; Physical Chemistry and Mass Spectrometry Laboratory
Fonds de la Recherche Fondamentale Collective - FRFC ; Fonds spéciaux, ULg ; Belgian program of Interuniversity Attraction Poles initiated by the Federal Office for Scientific Technical and Cultural Affaires ; Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/149136
10.1016/j.bbagen.2013.05.014

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