Purification and characterization of a mannose/N-acetyl-D-glucosamine-specific lectin from the seeds of Platymiscium floribundum Vogel |
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Authors: | Francisco Nascimento Pereira-Junior Helton Colares Silva Beatriz Tupinambá Freitas Bruno Anderson Matias Rocha Kyria Santiago Nascimento Celso Shinitti Nagano Rodrigo Bainy Leal Alexandre Holanda Sampaio Benildo Sousa Cavada |
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Institution: | Department of Biochemistry and Molecular Biology, Federal University of Ceara, Mister Hull s/n Bloco 907, 60.440-970, Fortaleza, CE, Brazil. |
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Abstract: | Platymiscium floribundum lectin (PFL), a mannose/N-acetyl-D-glucosamine-specific lectin, was isolated from P. floribundum seeds using Sepharose-mannose affinity media chromatography. PFL is a glycoprotein that is a potent agglutinin for rabbit erythrocytes. In addition, PFL is highly stable because it is able to maintain its hemagglutinating activity after exposure to temperatures of up to 60 °C for 1 h and exposure to a wide pH range. The PFL purification process was monitored using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the results showed that the purified lectin consists of a single band with a molecular mass of approximately 29 kDa in either the presence or the absence of a reducing agent. The analysis of purified PFL by electrospray ionization-mass spectrometry showed that most ions had a molecular weight of 27,053 ± 2 Da, and other less abundant ions had similar molecular weights. Gel filtration shows that the lectin exists as a dimer in solution with mass at approximately 65 kDa. Sixteen peptides were sequenced, and as a result, a total of 130 amino acids were identified and resulted in a coverage of approximately 65% of the PFL sequence. The partial sequence of PFL was aligned with sequences of other lectins from evolutionarily related species, and PFL showed considerable similarity to the other lectins. |
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Keywords: | plant lectin purification Platymiscium floribundum ESI mass spectrometry |
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