Detection of unwanted protein-bound ligands by capillary zone electrophoresis: the case of hidden ligands that stabilize cholinesterase conformation

Electrophoresis
D RochuP Masson

Abstract

Detection, identification and characterization of compounds present in purified proteins and biopharmaceuticals are of central interest. As well as chemical remedies, proteins of pharmacological interest have to exhibit their nakedness to become therapeutic drugs. Cholinesterases (ChE) are enzymes of major importance for detoxification of poisonous esters. Likewise, ChE are characterized by the high catalytic efficiency of an active site positioned at the bottom of a deep gorge. The gorge can be partially or fully occupied by ligands, i.e., substrates and inhibitors that are currently used in affinity chromatography purification steps. Accordingly, a suitable method allowing to analyse the presence of unwanted ligands and its influence on the functional conformation and stability of these enzymes was essential. We have developed CZE approaches for that purpose. The factors causing discrepancies between data for thermal unfolding of ChE by electrophoretic and by calorimetric methods were investigated. The presence of unwanted hidden ligands bound to purified enzymes was first demonstrated. The incidence of these ligands was discussed. Altogether, our results raised several questions concerning the real conformation of the native...Continue Reading

References

Nov 9, 1989·Biochimica Et Biophysica Acta·C S PayneA D Wolfe
Dec 1, 1994·Protein Science : a Publication of the Protein Society·J P Glusker
Jul 1, 1993·Progress in Neurobiology·J MassouliéF M Vallette
Oct 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·L IzadyarD Thomas
Oct 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·M HarelJ L Sussman
Jun 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·D R RipollJ L Sussman
Nov 1, 1995·Protein Science : a Publication of the Protein Society·D I KreimerL Weiner
Mar 20, 1999·Biochimica Et Biophysica Acta·J W TannerP A Liebman
May 18, 1999·Biochemical and Biophysical Research Communications·G Johnson, S W Moore

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Citations

Jun 1, 2007·Electrophoresis·Maria V PiaggioJulio A Deiber
May 24, 2006·Electrophoresis·Michael J LittlePieter K Roos
Nov 25, 2004·Journal of Chromatography. a·Hui XuHong-Yuan Chen

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