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Molecules 2011, 16(11), 9077-9089; https://doi.org/10.3390/molecules16119077

Molecular Characterization and Tandem Mass Spectrometry of the Lectin Extracted from the Seeds of Dioclea sclerocarpa Ducke

1
Laboratório de Moléculas Biologicamente Ativas (Biomol-Lab), Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Av. Humberto Monte s/n, Bloco 907, Lab. 1075, Campus do Pici, Fortaleza-CE, 60440-970, Brazil
2
Laboratório de Biotecnologia Molecular (LabBMol), Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Av. Humberto Monte s/n, Bloco 907, Lab. 1090, Campus do Pici, Fortaleza-CE, 60440-970, Brazil
3
Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, Santo Andre-SP, 09210-170, Brazil
4
Laboratório de Imunologia e Bioquímica de Sobral (LIBS), Faculdade de Medicina, Universidade Federal do Ceará, Sobral-CE, 62042-280, Brazil
5
Laboratório de Espectrometria de Massa Aplicado a Proteínas (LEMAP/Biomol-Lab), Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Av. Humberto Monte s/n, Bloco 825, Campus do Pici, Fortaleza-CE, 60440-970, Brazil
*
Author to whom correspondence should be addressed.
Received: 29 September 2011 / Revised: 20 October 2011 / Accepted: 21 October 2011 / Published: 28 October 2011
(This article belongs to the Section Molecular Diversity)
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Abstract

Lectin from the seeds of Dioclea sclerocarpa (DSL) was purified in a single step by affinity chromatography on a Sephadex G-50 column. The primary sequence, as determined by tandem mass spectrometry, revealed a protein with 237 amino acids and 81% of identity with ConA. DSL has a molecular mass of 25,606 Da. The β and γ chains weigh 12,873 Da and 12,752 Da, respectively. DSL hemagglutinated rabbit erythrocytes (both native and treated with proteolytic enzymes), showing stability even after one hour of exposure to a specific pH range. The hemagglutinating activity of DSL was optimal between pH 6.0 and 8.0, but was inhibited after incubation with D-galactose and D-glucose. The pure protein possesses a molecular mass of 25 kDa by SDS-PAGE and 25,606 Da by mass spectrometry. The secondary structure content was estimated using the software SELCON3. The results indicate that b-sheet secondary structures are predominant in DSL (approximately 42.3% antiparallel b-sheet and 6.7% parallel b-sheet). In addition to the b-sheet, the predicted secondary structure of DSL features 4.1% a-helices, 15.8% turns and 31.3% other contributions. Upon thermal denaturation, evaluated by measuring changes in ellipticity at 218 nm induced by a temperature increase from 20 °C to 98 °C, DSL displayed cooperative sigmoidal behavior with transition midpoint at 84 °C and permitted the observation of two-state model (native and denatured).
Keywords: plant lectin; diocleinae; Dioclea sclerocarpa plant lectin; diocleinae; Dioclea sclerocarpa
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Correia, J.L.A.; Nascimento, A.S.F.; Cajazeiras, J.B.; Gondim, A.C.S.; Pereira, R.I.; Sousa, B.L.; Silva, A.L.C.; Garcia, W.; Teixeira, E.H.; Nascimento, K.S.; Rocha, B.A.M.; Nagano, C.S.; Sampaio, A.H.; Cavada, B.S. Molecular Characterization and Tandem Mass Spectrometry of the Lectin Extracted from the Seeds of Dioclea sclerocarpa Ducke. Molecules 2011, 16, 9077-9089.

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