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Mediators of Inflammation
Volume 2005, Issue 5, Pages 263-272
http://dx.doi.org/10.1155/MI.2005.263

Characterization of Acetyl-CoA: Lyso-PAF Acetyltransferase of Human Mesangial Cells

1Faculty of Chemistry, National and Kapodistrian University of Athens, Panepistimioupolis, Athens 15771, Greece
2Centre for Nephrology, G. Papadakis General Hospital of Nikea-Pireaus, Athens 18454, Greece

Received 7 June 2005; Accepted 30 June 2005

Copyright © 2005 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Platelet activating factor (PAF) is a potent inflammatory mediator produced by various renal cells and it is implicated in renal pathology. The aim of this study is the characterization of remodeling lyso-PAF acetyltransferase, which is activated under inflammatory conditions, in human mesangial cell. Total membranes of mesangial cells were isolated and enzymatic activity and kinetic parameters were determined by trichloroacetic acid precipitation method. The effect of BSA, divalent cations, EDTA, and various chemicals on the activity of lyso-PAF acetyltransferase was also studied. Various detergents were also tested for the solubilization of the enzyme and only glycerol did not affect its activity. Partial purification of solubilized enzyme preparations of human kidney tissue and mesangial cells was performed on anion exchange column chromatography and native-PAGE electrophoresis and two active fractions were detected.