Premium
Purification and characterization of a mannose/ N ‐acetyl‐ d ‐glucosamine‐specific lectin from the seeds of Platymiscium floribundum Vogel
Author(s) -
PereiraJunior Francisco Nascimento,
Silva Helton Colares,
Freitas Beatriz Tupinambá,
Rocha Bruno Anderson Matias,
Nascimento Kyria Santiago,
Nagano Celso Shinitti,
Leal Rodrigo Bainy,
Sampaio Alexandre Holanda,
Cavada Benildo Sousa
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.2207
Subject(s) - lectin , molecular mass , mannose , glucosamine , size exclusion chromatography , biochemistry , dimer , sepharose , gel electrophoresis , polyacrylamide gel electrophoresis , mannan binding lectin , affinity chromatography , biology , chemistry , chromatography , enzyme , organic chemistry
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. Copyright © 2012 John Wiley & Sons, Ltd.