Properties of phospholipase A2 activity from bovine retinal rod outer segments

Experimental Eye Research
P I Castagnet, Norma M Giusto

Abstract

In the present paper the properties of a phospholipase A2 (PLA2) activity associated with rod outer segments (ROS) have been studied. Under adequate experimental conditions, ROS PLA2 activity presented a maximum at pH 9.0. When using 1-palmitoyl-2[1- 14C]arachidonoyl-sn-glycero-3-phosphocholine as substrate, the apparent Km value was 36.5 microM and the Vmax value was 0.612 nanomol per hour per mg protein. The enzyme was fully activated at free calcium concentrations in the range 100- 300 microM. Concentrations of CaCl2 above 1 milliM inhibited its activity as a function of the ion concentration. The presence of EGTA or EDTA caused a 73% inhibition of PLA2 activity in ROS relative to the activity observed when no calcium was added. Treatment of the membranes with different kinds of detergents (Triton X-100, sodium deoxycholate and CHAPS) at concentrations below and above their critical micelle concentration resulted in an inhibition of PLA2 activity. However, when Triton X-100 was present at a concentration of 0.05 milliM, no significant change in enzymatic activity could be observed. Maximum inhibition was observed in the presence of CHAPS 25 milliM (87%). Seventy-five percent of PLA2 activity was recovered in ROS membranes af...Continue Reading

Citations

Oct 16, 2009·Journal of Lipid Research·Norma M GiustoMónica G Ilincheta de Boschero
Apr 15, 2011·Journal of Lipids·Susana J PasquaréNorma M Giusto
Apr 20, 2017·Frontiers in Pharmacology·Carmelina D AnfusoClaudio Bucolo
Jul 21, 2004·Journal of Neuroscience Research·Miriam KolkoNicolas G Bazan
May 10, 2007·Acta Ophthalmologica Scandinavica·Miriam KolkoSteffen Heegaard
Aug 9, 1996·The Journal of Biological Chemistry·M JacobC Salesse
Jul 23, 1999·Current Eye Research·S D Wilt, B J McLaughlin
Jun 17, 2020·Journal of Lipid Research·Raju V S Rajala
Feb 9, 2021·Journal of Lipid Research·Raju V S Rajala
Jul 19, 2000·Archives of Biochemistry and Biophysics·S J PasquaréN M Giusto
Apr 1, 1997·Archives of Biochemistry and Biophysics·P I Castagnet, N M Giusto
Dec 17, 1998·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·P I CastagnetN M Giusto
Jun 17, 2000·Progress in Lipid Research·N M GiustoM G Ilincheta de Boschero
Jun 14, 2002·Archives of Biochemistry and Biophysics·P I Castagnet, N M Giusto
Apr 27, 2005·Prostaglandins, Leukotrienes, and Essential Fatty Acids·Pierre HardySylvain Chemtob
Oct 18, 2008·Neurochemistry International·S J Pasquaré, N M Giusto
Apr 21, 2010·Biochimie·Jinmei Wang, Miriam Kolko

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