Biochemical characterization of a cysteine proteinase from Bauhinia forficata leaves and its kininogenase activity

Biochemical characterization of a cysteine proteinase from Bauhinia forficata leaves and its kininogenase activity

Author Andrade, Sheila Siqueira Autor UNIFESP Google Scholar
Silva-Lucca, Rosemeire Aparecida Google Scholar
Santana, Lucimeire Aparecida de Autor UNIFESP Google Scholar
Gouvea, Iuri Estrada Autor UNIFESP Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Carmona, Adriana Karaoglanovic Autor UNIFESP Google Scholar
Araujo, Mariana da Silva Autor UNIFESP Google Scholar
Sampaio, Misako Uemura Autor UNIFESP Google Scholar
Oliva, Maria Luiza Vilela Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Estadual Oeste Parana
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.
Keywords Circular dichroism
Cysteine proteinase
Plant enzyme
Protein purification
Substrate specificity
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Date 2011-02-01
Published in Process Biochemistry. Oxford: Elsevier B.V., v. 46, n. 2, p. 572-578, 2011.
ISSN 1359-5113 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 572-578
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000287274000021

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