PO41, the anti-hemorragina present in the serum of Philander opossum: isolation, characterization and interaction with metalloproteases isolated of the poison of Bothrops jararaca
An antihaemorrhagic factor from Philander opossum serumwas isolated and characterizated. In order to study theneutralization effect of this inhibitor, we have developed a new methodology to isolate a PI as well as PIII SVMP from Bothrops jararaca venom (Bjv). The antihaemorrhagic protein was isolated from Philander opossum serum by DEAE-Sephacel, Phenyl Superose and Superdex 200 hromatographies. The homogenous proteinpresented a molecular mass of 41,330 amu by MALDI-TOF mass spectrometry and was named PO41. By gel filtrationchromatography, we have obtained a molecular mass of 61,5 kDa on native conditions. From Edman sequencing, PO41 and DM43, another SVMP inhibitor, presented similar amino termini through the first 27 residues. Identical immunoblotting patterns were obtained when both inhibitors were developed with polyclonal antibodies raised against Didelphis marsupialis active serum fraction. Isoeletric points were estimated to be less than 3.5 for both proteins. The new methodology to purify jararhagin (a PIII SVMP) and botrolysin (a PI SVMP) was based on cromatographies using Superdex 200 and Phenyl Superose. Jarahagin was further submitted to a Mono Q column. The proteolytic and haemorrhagic effects of the haemorrhagins were neutralized by PO41. Both haemorrhagins formed stable complexes with PO41, as observed by gel filtration on Superdex 200. The results suggest that PO41 presents similar physicochemical, structural, immunoreactive and biological properties with the SVMP inhibitors, including DM40 e DM43. These inhibitors belong to the supergene family of immunoglobulins.
Advisor:Gilberto Barbosa Domont; Jonas Enrique Aguilar Perales
School:Faculdades Oswaldo Cruz
Source Type:Master's Thesis
Keywords:Philander opossum metaloproteases Bothrops jararaca metalloproteases
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