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

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; in revised form: 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

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MDPI and ACS Style

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.

AMA Style

Correia JLA, Nascimento ASF, Cajazeiras JB, Gondim ACS, Pereira RI, Sousa BL, Silva ALC, Garcia W, Teixeira EH, Nascimento KS, Rocha BAM, Nagano CS, Sampaio AH, Cavada BS. Molecular Characterization and Tandem Mass Spectrometry of the Lectin Extracted from the Seeds of Dioclea sclerocarpa Ducke. Molecules. 2011; 16(11):9077-9089.

Chicago/Turabian Style

Correia, Jorge Luis Almeida; Nascimento, Antônia Sâmia Fernandes do; Cajazeiras, João Batista; Gondim, Ana Cláudia Silva; Pereira, Ronniery Ilario; Sousa, Bruno Lopes de; Silva, André Luiz Coelho da; Garcia, Wanius; Teixeira, Edson Holanda; Nascimento, Kyria Santiago do; Rocha, Bruno Anderson Matias da; Nagano, Celso Shiniti; Sampaio, Alexandre Holanda; Cavada, Benildo Sousa. 2011. "Molecular Characterization and Tandem Mass Spectrometry of the Lectin Extracted from the Seeds of Dioclea sclerocarpa Ducke." Molecules 16, no. 11: 9077-9089.

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