NDLI: Isolation of an Entamoeba histolytica intracellular alkaline phospholipase A2

Content Provider	Springer Nature Link
Author	Vargas Villarreal, J. Olvera Rodríguez, A. Mata Cárdenas, B. D. Martínez Rodríguez, H. G. Said Fernández, S. Alagón Ca, A.
Copyright Year	1998
Abstract	The major hemolytic activity of Entamoeba histolytica is located in a subcellular fraction called P30. Its maximal effect is observed at pH 8.0 and 1 mM Ca2+ and is due to a phospholipase A (PLA). In the present study a membrane-associated phospholipase A2 was purified from P30 to homogeneity. P30 was fractionated with ethyl ether and the insoluble fraction was extracted with 1 M KCl. The KCl-soluble material was diluted ten times with 0.1 M TRIS-HCl (pH 9.5) and passed through a chromatofocusing column with a 9–4 pH gradient. Four peaks with PLA2 activity were obtained. By affinity chromatography, peak II, the one with the highest specific activity, was resolved in three more PLA2 peaks. Peak II.2 had the highest PLA2 specific activity. When analyzed by sodium dodecyl sulfate-polyacrylamide slab-gel electrophoresis under nonreducing conditions, peak II.2 yielded a single band with an apparent molecular mass of 30 kDa. Under reducing conditions the protein dissociated into two 15-kDa monomers. The purified PLA II.2 displayed its activity at the same conditions under which the P30 hemolytic activity was maximal. The isoelectric point of PLA II.2 was 7.0. The purification procedure described above provides sufficient material for determination of the relative importance of the enzyme in the E. histolytica pathogenic mechanisms.
Starting Page	310
Ending Page	314
Page Count	5
File Format	PDF
ISSN	09320113
Journal	Parasitology Research
Volume Number	84
Issue Number	4
e-ISSN	14321955
Language	English
Publisher	Springer-Verlag
Publisher Date	1998-03-23
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Veterinary Parasitology Insect Science

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