PMID: 3754767Jun 13, 1986Paper

Contamination of commercial preparations of xanthine oxidase by a Ca2+-dependent phospholipase A2

Biochimica Et Biophysica Acta
D A GamacheR C Franson

Abstract

Using [1-14C]oleate-labelled autoclaved Escherichia coli as substrate, we demonstrate that many, but not all, commercial preparations of xanthine oxidase contain phospholipase A2 activity as a contaminant. Phospholipase A2 activity (64.3-545.6 nmol phospholipid hydrolyzed per min per mg protein) was optimal in the neutral to alkaline pH range, was Ca2+-dependent, and was unaffected by the addition of xanthine. Phospholipase A2 activity was totally inhibited by 1.0 mM EDTA while radical production by xanthine plus xanthine oxidase was unaffected by EDTA. Even chromatographically purified xanthine oxidase (Sigma Grade III) contained substantial phospholipase A2 activity (64.3 nmol/min per mg). Since the preparation of xanthine oxidase employs proteolytic digestion of milk or buttermilk by pancreatin, an extract of pancreas which is an organ rich in phospholipase A2 activity, we speculate that the contaminant phospholipase A2 is introduced by this treatment. Because xanthine oxidase is used extensively to study free radical-induced cell injury and membrane phospholipid alterations, the presence of a potent extracellular phospholipase A2 may have influenced previously published reports and such studies in the future should be inter...Continue Reading

References

Dec 18, 1979·Biochimica Et Biophysica Acta·R L Jesse, R C Franson
Jan 1, 1976·Annual Review of Pharmacology and Toxicology·G L Plaa, H Witschi
Aug 1, 1975·Archives of Biochemistry and Biophysics·W R WaudK V Rajagopalan
Apr 24, 1968·Biochimica Et Biophysica Acta·G H de HaasL L van Deenen
Aug 1, 1981·Journal of Molecular and Cellular Cardiology·M L HessH A Kontos
Jun 1, 1982·Archives of Biochemistry and Biophysics·M TienS D Aust
Sep 15, 1984·The Biochemical Journal·A Ager, J L Gordon
Oct 15, 1984·Biochemical and Biophysical Research Communications·W B WeglickiI T Mak

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Citations

Sep 15, 1987·Biochemical and Biophysical Research Communications·G Thomas, P W Ramwell
Mar 13, 1987·Biochemical and Biophysical Research Communications·A K Grover, S E Samson
Jan 19, 1988·Biochimica Et Biophysica Acta·D A GamacheR C Franson

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