Purification and primary structure of a novel mannose-specific lectin from Centrolobium microchaete Mart seeds

Abstract

This study aimed to purify and characterize a novel mannose-binding lectin from the seeds of Centrolobium microchaete. Centrolobium microchaete lectin (CML) was purified by affinity chromatography in mannose-Sepharose-4B column. CML agglutinated rabbit erythrocytes and was inhibited by D-mannose, α-methyl-D-mannoside, D-glucose, N-Acetyl-D-glucosamine and sucrose. The lectin was stable at pH 7.0 and 8.0 and temperatures up to 60°C. The monomeric form of CML showed approximately 28kDa, and its native form is probably a homodimer, as determined by gel filtration chromatography. The primary structure of CML was determined by tandem mass spectrometry that showed CML as a protein with two distinct forms (isolectins CML-1 and CML-2) with 246 and 247 residues, respectively. CML-2 possesses one residue of Asn more than CML-1 in C-terminal. The primary structure of CML agrees with the molecular weights found by electrospray ionization mass spectrometry: 27,224 and 27,338Da for CML-1 and CML-2, respectively. CML is a metal-dependent glycoprotein. Moreover, the glycan composition of CML and its structure were predicted.

In-vitro Antioxidant and Antibacterial Potential of Mannose/Glucose-binding Pterocarpus osun Craib. Seeds Lectin

“…The results are shown in Table 2. Similar results were reported for lectins from Platymiscium floribundum [16], Centrolobium microchaete [18] and Canavalia virosa [37]. Lis and Sharon [38] revealed that trypsin can be used to modify the erythrocytes surface to enhance its affinity for lectins without affecting the total number of lectin binding sites on the erythrocytes.…”

“…POSL belongs to the mannose/glucose specificity group of lectins from Dalbergieae tribe, which has specificity for different sugars. Among well studied member of the tribe that belong to mannose/glucose specificity group are lectins from Pterocarpus angolensis [3], Platymiscium floribundum [16], Centrolobium microchaete [18], and Platypodium elegans [40]. Though, other members of the tribe that have specificity for other carbohydrates especially galactose have also been reported (Vatairea marcocarpa [11]; Lonchocarpus capassa [14]; Vatairea guianensis [15]).…”

“…Other seed lectins that have been purified and biochemically characterized from this tribe include galactosebinding lectins from Lonchocarpus capassa [14], Vatairea macrocarpa [11] and Vatairea guianensis [15] and mannose-binding lectins from Platymiscium floribundum [16] and Centrolobium tomentosum [17]. Amino acid sequence of P. floribundum [16], C. tomentosum [17] and Centrolobium microchate [18] lectins has been determined partially. Generally, legume lectins are structurally homologous and at time have similar physicochemical properties but display biological activities that are distinctly differ.…”

Biochemical characterisation of lectin from Indian hyacinth plant bulbs with potential inhibitory action against human cancer cells

“…Carbohydrate content and glycosylation nature of Hyacinth bulb lectin was determined by anthrone assay and PAS staining respec- tively. The carbohydrate content of purified lectin was found to be 5.2% and according to literature carbohydrate content of lectin vary from 1.5 to 16% [33,34]. PAS stain result indicated that hyacinth lectin was glycoprotein in nature ( Fig.…”

Plant Lectins with Insecticidal and Insectistatic Activities

“…For these lectins, SDS-PAGE profile was similar to other mannose lectins, a band around 30 kDa with an isoelectric point near to 6.5, and they were able to agglutinate human RBCs from A, B, and O donors. This means that specificity by mannose/glucose moieties or mannose-rich glycan is not a unique feature of any family; conversely, species such as Galanthus nivalis (tribe Galantheae) [105] and Centrolobium microchaete (tribe Dalbergieae) [106], among others, even species from other families such as Moraceae have mannose/glucose lectins [107]. Galactia lindenii lectin type -I (GLL-I) [77].…”