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Purification and partial characterization of a novel lectin from Dioclea lasiocarpa Mart seeds with vasodilator effects
Author(s) -
do Nascimento Antônia Sâmia F.,
Gondim Ana Cláudia S.,
Cajazeiras João B.,
Correia Jorge Luis A.,
Pires Alana de F.,
do Nascimento Kyria S.,
Silva André Luis C.,
Nagano Celso S.,
Assreuy Ana Maria S.,
Cavada Benildo S.
Publication year - 2012
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.2222
Subject(s) - chromatography , chemistry , lectin , affinity chromatography , phenylephrine , sephadex , molecular mass , biochemistry , biology , enzyme , endocrinology , blood pressure
A lectin from seeds of Dioclea lasiocarpa (DLL) was purified in a single step by affinity chromatography in a Sephadex G‐50 column. DLL haemagglutinated rabbit erythrocytes showing stability even after 1 h of exposure to a different pH values (optimal between pH 6.0 and 8.0) but was inhibited after incubation with d ‐mannose and d ‐glucose. The pure protein possessed a molecular weight of 25 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 25,410Da by mass spectrometry. The results analyzed by the software SELCON 3 indicate that β ‐sheet secondary structures are predominant in DLL (approximately 40.2% antiparallel β ‐sheet, 4.6% parallel β ‐sheet, 7.2% α ‐helices, 17.3% turns, and 28.7% unordered structures). Mechanical activity of isolated aorta from rat measured by cumulative concentration curves of DLL, performed at the contraction plateau induced by phenylephrine in either endothelium‐intact or denuded aorta. DLL (IC 50 = 34.12 ± 3.46 µg/ml) relaxed precontracted endothelized aortic rings by 34.61 ± 9.06%, 55.19 ± 11.9%, and 81.33 ± 14.35%, respectively, at 10 µg/ml (initial concentration), 30 µg/ml, and 100 µg/ml (maximum effect). All effects occurred via interaction with lectin domains and participation of nitric oxide. Copyright © 2012 John Wiley & Sons, Ltd.