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Báo cáo Y học: Galactosyl-mimodye ligands for Pseudomonas fluorescens b-galactose dehydrogenase

Protein molecular modelling and ligand docking were employed for the design of anthraquinone galactosyl-bio-mimetic dye ligands (galactosyl-mimodyes) for the target enzyme galactose dehydrogenase (GaDH). Using appro-priate modelling methodology, a GaDH model was build based on a glucose-fructose oxidoreductase (GFO) protein template. Subsequent computational analysis predicted chimaeric mimodye-ligands comprising a NAD-pseudomi-metic moiety (anthraquinone diaminobenzosulfonic acid) and a galactosyl-mimeticmoiety (2-amino-2-deoxygalactose or shikimic acid) bearing an aliphatic linker molecule. . | Eur. J. Biochem. 269 5391-5405 2002 FEBS 2002 doi 10.1046 j.1432-1033.2002.03211.x Galactosyl-mimodye ligands for Pseudomonas fluorescens b-galactose dehydrogenase Design synthesis and evaluation C. F. Mazitsos1 D. J. Rigden2 P. G. Tsoungas3 and Y. D. Clonis1 1 Laboratory of Enzyme Technology Department of Agricultural Biotechnology Agricultural University of Athens Greece 2Embrapa Recursos Geneticos e Biotecnologia Brasilia Brazil 3Department of Pharmaceutical and Biological Chemistry School of Pharmacy University of London UK Protein molecular modelling and ligand docking were employed for the design of anthraquinone galactosyl-biomimetic dye ligands galactosyl-mimodyes for the target enzyme galactose dehydrogenase GaDH . Using appropriate modelling methodology a GaDH model was build based on a glucose-fructose oxidoreductase GFO protein template. Subsequent computational analysis predicted chimaeric mimodye-ligands comprising a NAD-pseudomi-metic moiety anthraquinone diaminobenzosulfonic acid and a galactosyl-mimetic moiety 2-amino-2-deoxygalactose or shikimic acid bearing an aliphatic linker molecule. In addition the designed mimodye ligands had an appropriate in length and chemical nature spacer molecule via which they can be attached onto a chromatographic support without steric clashes upon interaction with GaDH. Following their synthesis purification and analysis the ligands were immobilized to agarose. The respective affinity adsorbents compared to other conventional adsorbents were shown to be superior affinity chromatography materials for the target enzyme Pseudomonas fluorescens b-galactose dehydrogenase. In addition these mimodye affinity adsorbents displayed good selectivity binding low amounts of enzymes other than GaDH. Further immobilized dye-ligands comprising different linker and or spacer molecules or not having a biomimetic moiety had inferior chromatographic behavior. Therefore these new mimodyes suggested by computational analysis are .