Multiple advantages of capillary zone electrophoresis for exploring protein conformational stability

Abstract

This review summarizes the work of our laboratory to explore the use of capillary zone electrophoretic (CZE) methods for the investigation of protein conformational stability. Early CZE works on protein denaturation as well as fundamental and theoretical considerations are discussed. Instrumental aspects of the CE-based approach including general and particular CE requirements are documented. Several aspects dealing with estimation of stability of enzymes (cholinesterases and organophosphate-hydrolyzing enzymes) interacting with organophosphates profusely illustrate the multiple advantages of CZE. The discrimination of parameters controlling the "good compromise" stability/plasticity for allowing functional efficiency of these enzymes is exemplified. Thermal stability, susceptibility to high electric field, alteration of stability by bound ligands and the role of associated cations in metalloenzymes have been successfully investigated.

Analysis of recombinant human growth hormone by capillary electrophoresis with bilayer-coated capillaries using UV and MS detection

“…Capillary electrophoresis (CE) is a powerful separation tool, which in principle is very well suited for the analysis of proteins [5][6][7][8]. CE analyses are relatively fast and separation efficiencies can be quite high.…”

Subject Index To Volume 2

“…Protein unfolding studies by CE of phosphotriesterase (PTE) with various metal cations substituted into the active site [24,25] as well as different cho-linesterases [21][22][23] have been reviewed by Rochu and Masson [31]. The results obtained by thermal unfolding studies were validated using differential scanning calorimetry.…”

“…In this case, receptor-ligand binding results in a significant enhancement in protein conformational stability relative to the ligand-free receptor. Since there have been several reviews on protein unfolding by CE published between 1995 and 2002 [14,[30][31], the focus of this review will highlight the general theory and application of protein unfolding and conformational studies in CE with emphasis on recent developments relevant to holoprotein and protein aggregation systems. Basic models and assumptions used to interpret mobility changes in CE are also presented, including relevant thermodynamic and kinetic aspects of protein unfolding.…”

Determination of small phosphorus-containing compounds by capillary electrophoresis

“…2a). Rochu and Masson summarizes the use of CZE methods for the investigation of the interactions between proteins and OP [47]. Two different soil bacteria, Pseudomonas diminuta and Flavobacterium sp.…”