Biomimetic dyes as affinity chromatography tools in enzyme purification.
Affinity adsorbents based on immobilized triazine dyes offer important advantages circumventing many of the problems associated with biological ligands. The main drawback of dyes is their moderate selectivity for proteins. Rational attempts to tackle this problem are realized through the biomimetic dye concept according to which new dyes, the biomimetic dyes, are designed to mimic natural ligands. Biomimetic dyes are expected to exhibit increased affinity and purifying ability for the targeted proteins. Biocomputing offers a powerful approach to biomimetic ligand design. The successful exploitation of contemporary computational techniques in molecular design requires the knowledge of the three-dimensional structure of the target protein, or at least, the amino acid sequence of the target protein and the three-dimensional structure of a highly homologous protein. From such information one can then design, on a graphics workstation, the model of the protein and also a number of suitable synthetic ligands which mimic natural biological ligands of the protein. There are several examples of enzyme purifications (trypsin, urokinase, kallikrein, alkaline phosphatase, malate dehydrogenase, formate dehydrogenase, oxaloacetate decarboxylase and lactate dehydrogenase) where synthetic biomimetic dyes have been used successfully as affinity chromatography tools.
Clonis
Y D
YD
Department of Agricultural Biotechnology, Agricultural University of Athens, Greece. clonis@aua.gr
Labrou
N E
NE
Kotsira
V P
VP
Mazitsos
C
C
Melissis
S
S
Gogolas
G
G
D002846:Chromatography, Affinity / Q000379:methods*; D004396:Coloring Agents / Q000737:chemistry*; D004798:Enzymes / Q000302:isolation & purification*; D018716:Molecular Mimicry
D004396:Coloring Agents; D004798:Enzymes