Characterization of acetyl-CoA: lyso-PAF acetyltransferase of human mesangial cells.
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.
Fragopoulou
Elizabeth
E
Faculty of Chemistry, National and Kapodistrian University of Athens, and Centre for Nephrology, G. Papadakis General Hospital of Nikea-Pireaus, Greece.
Iatrou
Christos
C
Demopoulos
Constantinos Alexandros
CA
D000123:Acetyltransferases / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000166:cytology; D050527:Mesangial Cells / Q000737:chemistry / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000096:biosynthesis*
D010972:Platelet Activating Factor; D000123:Acetyltransferases; C046035:1-alkylglycerophosphocholine acetyltransferase