Purification, Partial Characterization and Immobilization of a Mannose-Specific Lectin from Seeds of Dioclea lasiophylla Mart

Molecules

Vanir Reis Pinto Júnior³  Mayara Queiroz de Santiago³  Vinໜius José da Silva Osterne³  Jorge Luis Almeida Correia³  Francisco Nascimento Pereira Júnior³  João Batista Cajazeiras³  Mayron Alves de Vasconcelos³  Edson Holanda Teixeira²  Antônia Sâmia Fernandes do Nascimento³  Thaiz Batista Azevedo Rangel Miguel³  Emilio de Castro Miguel³  Alexandre Holanda Sampaio¹  Kyria Santiago do Nascimento³  Celso Shiniti Nagano¹ 

[1] Laboratory of Mass Spectrometry Applied to Proteins (LEMAP), Federal University of Ceará, Av. Humberto Monte s/n, Bloco 825, Campus do Pici, Fortaleza-CE 60440-970, Brazil; E-Mails:;Integrated Laboratory of Biomolecules (LIBS), Federal University of Ceará, Department of Pathology and Legal Medicine, Faculty of Medicine, Fortaleza, CE 60430-160, Brazil; E-Mail:;Laboratory of Biologically Active Molecules (Biomol-Lab), Department of Biochemistry and Molecular Biology, Federal University of Ceará, Av. Humberto Monte s/n, Bloco 907, Lab. 1075, Campus do Pici, Fortaleza-CE 60440-970, Brazil; E-Mails:

关键词: lectin;    Dioclea lasiophylla;    Diocleinae;    toxicity;    immobilization;   

DOI  :  10.3390/molecules180910857

来源: mdpi

【 摘 要 】

Lectin from the seeds of Dioclea lasiophylla (DlyL) was purified in a single step by affinity chromatography on a Sephadex^® G-50 column. DlyL strongly agglutinated rabbit erythrocytes and was inhibited by monosaccharides (_D-mannose and α-methyl-d-mannoside) and glycoproteins (ovalbumin and fetuin). Similar to other Diocleinae lectins, DlyL has three chains, α, β and γ, with mass of 25,569 ± 2, 12,998 ± 1 and 12,588 ± 1 Da, respectively, and has no disulfide bonds. The hemagglutinating activity of DlyL was optimal in pH 8.0, stable at a temperature of 70 °C and decreased in EDTA solution, indicating that lectin activity is dependent on divalent metals. DlyL exhibited low toxicity on Artemia sp. nauplii, but this effect was dependent on the concentration of lectin in solution. DlyL immobilized on cyanogen bromide-activated Sepharose^® 4B bound 0.917 mg of ovalbumin per cycle, showing the ability to become a tool for glycoproteomics studies.

【 授权许可】

CC BY   
© 2013 by the authors; licensee MDPI, Basel, Switzerland.