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Efficient Immobilization of Phospholipase A2

  • Wonhwa Cho
  • Zhen Shen
Part of the Methods in Molecular Biology™ book series (MIMB, volume 109)

Abstract

Immobilized enzymes have been widely used for biomedical and industrial applications. Since phospholipase A2 (PLA2; E.C.3.1.1.4) catalyzes the hydrolysis of a wide range of phospholipid aggregates, immobilized PLA2 should also have potential biomedical applications. For instance, a novel therapy for hypercholesterolemia has been recently developed that utilizes immobilized PLA2 contained in an extracorporeal shunt, based on the finding that human serum low density lipoprotein modified by immobilized PLA2 was removed from the circulation by liver more rapidly than unmodified low density lipoprotein (1). Also, immobilized PLA2 has been used to study the mechanism of interfacial activation of PLA2 (2,3). To date, the immobilization of PLA2 has been carried out primarily with cobra venom enzymes using two different coupling methods; the coupling through the ɛ-amino group of lysines and the coupling through the carboxyl groups of asparatates and glutamates (3). These immobilization methods, however, often lead to a significant loss of enzymatic activity of PLA2, thereby limiting the application of immobilized PLA2. For instance, when the ɛ-amino groups are coupled to a solid support, the coupling yield is high but the activity of immobilized enzyme toward aggregated phospholipid substrates decreases drastically, which has been ascribed to the modification of the lysyl residues that play critical roles in the interaction of PLA2s with anionic interfaces. On the other hand, the coupling of protein carboxylic groups to the solid support is hampered by a lower coupling yield and modification of catalytically important residues. On the other hand, the coupling of protein carboxylic groups to the solid support is hampered by a lower coupling yield and modification of catalytically important residues.

Keywords

High Performance Liquid Chromatography Coupling Yield Octanoic Acid Immobilization Method Aqueous Acetic Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc, Totowa, NJ 1999

Authors and Affiliations

  • Wonhwa Cho
    • 1
  • Zhen Shen
    • 2
  1. 1.Department of ChemistryUniversity of Illinois at ChicagoChicago
  2. 2.Department of ChemistryUniversity of IllinoisChicago

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