STUDIES ON PHOSPHOLIPASE A2 (PLA2) INHIBITOR FROM Echis ocellatus SERUM AND ITS EFFECT ON NEUROTOXIC PLA2 AND NON TOXIC SECRETORY PLA2

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dc.contributor.authorADAMUDE, Fatima Amin
dc.date.accessioned2015-03-16T09:32:44Z
dc.date.available2015-03-16T09:32:44Z
dc.date.issued2014-08
dc.descriptionA THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY ZARIA, NIGERIA, IN PARTIAL FULFILMENT FOR THE AWARD OF MASTERS OF SCIENCE DEGREE IN BIOCHEMISTRY DEPARTMENT OF BIOCHEMISTRY FACULTY OF SCIENCE AHMADU BELLO UNIVERSITY, ZARIA AUGUST, 2014en_US
dc.description.abstractThe most effective and acceptable therapy for snakebite victims is the immediate administration of antivenin which is limited by problems of hypersensitivity reactions in some individuals and its inability to resolve the local effects of the venom. The aim of this study was to investigate the presence and specificity of Phospholipase A2 Inhibitor from Echis ocellatus serum towards snake venom neurotoxic PLA2 and mammalian secretory PLA2. Phospholipase A2 (EC.3.1.1.4) was isolated and partially purified from the venom of Echis ocellatus (carpet viper). A two step purification process on DEAE-cellulose and Sephadex G-75 column chromatography gave a specific activity of 53.17μmol/min/mg protein, 16.36 purification fold and 43.11% recovery. Initial velocity studies for the determination of kinetic constants using L-α- Lecithin as substrate revealed a KM and Vmax of 1.4mgml-1 and 4.5μmolmin-1 respectively. A protein (Phospholipase A2 Inhibitor from Echis ocellatus Serum (PIES)) isolated and purified from E. ocellatus serum inhibited the carpet viper PLA2 enzyme in a dose dependent manner. A two step purification process on Sephadex G-200 column and DEAE- cellulose chromatography gave 2 active fractions that inhibited the venom PLA2 by 78%. The result from SDS-PAGE showed the inhibitor to be a 24.98kDa protein and its kinetic study revealed a mixed pattern of inhibition on the carpet viper PLA2 with an estimated Ki values of 3.8%(v/v) to 7.3%(v/v). Membrane stabilizing and protective ability of PIES was recorded by its potential to reduce hemolysis due to venom PLA2 from 81.20 % to 35.30 % in vitro. Coagulant potentials of PIES were also seen in its ability to restore plasma coagulation time to less than a minute. Interestingly, PIES does not affect the enzymatic activity of mammalian secretory PLA2 but strongly inhibits PLA2 activity of carpet viper in this study. In conclusion, the present study shows that PIES may be a promising candidate for future development of a novel antivenin drug.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/6219
dc.language.isoenen_US
dc.subjectSTUDIES,en_US
dc.subjectPHOSPHOLIPASE A2en_US
dc.subject(PLA2) INHIBITOR,en_US
dc.subjectchis ocellatus,en_US
dc.subjectSERUM,en_US
dc.subjectEFFECT,en_US
dc.subjectNEUROTOXIC PLA2,en_US
dc.subjectNON TOXIC,en_US
dc.subjectSECRETORY PLA2en_US
dc.titleSTUDIES ON PHOSPHOLIPASE A2 (PLA2) INHIBITOR FROM Echis ocellatus SERUM AND ITS EFFECT ON NEUROTOXIC PLA2 AND NON TOXIC SECRETORY PLA2en_US
dc.typeThesisen_US
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