Artículos de revistas
Enzymatic And Structural Characterization Of New Pla2 Isoform Isolated From White Venom Of Crotalus Durissus Ruruima
Registro en:
Toxicon. , v. 53, n. 1, p. 104 - 114, 2009.
410101
10.1016/j.toxicon.2008.10.021
2-s2.0-57849150819
Autor
Diz Filho E.B.S.
Marangoni S.
Toyama D.O.
Fagundes F.H.R.
Oliveira S.C.B.
Fonseca F.V.
Calgarotto A.K.
Joazeiro P.P.
Toyama M.H.
Institución
Resumen
This work reports the structural and enzymatic characterization of a new sPLA2 from the white venom of Crotalus durissus ruruima, nominated PLA2A. The homogeneity of the PLA2A fraction and its molecular mass were initially evaluated by SDS-PAGE and confirmed by MALDI-TOF spectrometry, indicating a molecular mass of 14,299.34 Da. Structural investigation, through circular dichroism spectroscopy, revealed that PLA2A has a high content of alpha helix and beta-turn structures, 45.7% and 35.6% respectively. Its amino acid sequence, determined by Edman degradation and "de novo amino acid sequencing", exhibited high identity to PLA2 Cdt F15 from Crotalus durissus terrificus. The enzymatic investigation, conducted using the synthetic substrate 4-nitro-3-(octanoyloxy)-benzoic acid, determined its Vmax (7.56 nmoles/min) and KM (2.76 mM). Moreover, PLA2A showed an allosteric behavior and its enzymatic activity was dependent on Ca2+. 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