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Title: Biochemical characterization of a cysteine proteinase from Bauhinia forficata leaves and its kininogenase activity
Authors: Andrade, Sheila Siqueira [UNIFESP]
Silva-Lucca, Rosemeire Aparecida
Santana, Lucimeire Aparecida de [UNIFESP]
Gouvea, Iuri Estrada [UNIFESP]
Juliano, Maria Aparecida [UNIFESP]
Carmona, Adriana Karaoglanovic [UNIFESP]
Araujo, Mariana da Silva [UNIFESP]
Sampaio, Misako Uemura [UNIFESP]
Oliva, Maria Luiza Vilela [UNIFESP]
Keywords: Circular dichroism
Cysteine proteinase
Plant enzyme
Protein purification
Substrate specificity
Issue Date: 2011-02-01
Publisher: Elsevier B.V.
Citation: Process Biochemistry. Oxford: Elsevier B.V., v. 46, n. 2, p. 572-578, 2011.
Abstract: In this work, the proteinase activity detect in the acetone precipitate (80%, v/v) of B. forficata leaves, trivially known as cow paw, and popularly used in folk medicine for treatment of diabetes mellitus, was purified by chromatography on Sephadex G-25, Canecystatin-Sepharose, and on Con A-Sepharose. the molecular weight 30 kDa was estimated by SDS-PAGE and zymography, and the N-terminal sequence and CD spectra indicated a relationship with the papain family of cysteine proteinases. Denominated baupain, the enzyme was activated by dithiotreitol and inhibited by E-64 and iodoacetamide, but not by benzamidine, TLCK. TPCK and EDTA. the 52 and S1 substrate specificity of baupain, assayed with two series of fluorescence resonance energy transfer (FRET) peptide substrates derived from Abz-KLRSSK-Q-EDDnp, indicates a preference for Phe and Tyr at P2 position over Leu found in papain. Baupain releases bradykinin from HMWK (human high molecular weight kininogen) though its proteolytic activity is blocked by the sequence motif QVVA of kininogen (K(iapp) = 1.9 x 10(-8) M). Canecystatin, from sugar cane, which also lodges the QVVA sequence, inhibits baupain (K(iapp) = 0.18 x 10(-9) M). (C) 2010 Elsevier B.V. All rights reserved.
metadata.dc.language.iso: eng
Other Identifiers:
metadata.dc.rights: Acesso aberto
metadata.dc.type: Artigo
metadata.dc.format.extent: 572-578
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