A Diocleinae type II lectin from Dioclea lasiophylla Mart. Ex Benth seeds specific to α-lactose/GalNAc. (June 2020)

• Record Type: Journal Article
• Authors: Cavada, Benildo Sousa; Pinto-Junior, Vanir Reis; Osterne, Vinicius Jose Silva; Lossio, Claudia Figueiredo; Silva, Mayara Torquato Lima; Correia, Jorge Luis Almeida; Correia, Sarah Elizabeth Gomes; Nagano, Celso Shiniti; Oliveira, Messias Vital; Lima, Lara Dias; Vital, Ana Paula Moreira Sousa; Leal, Rodrigo Bainy; Nascimento, Kyria Santiago
• Is Part Of: Process biochemistry. Volume 93(2020)
• Page Start: 104
• Page End: 114
• Publication Date: 2020-06
• Subjects: ConA Concanavalin A -- DLL2 Dioclea lehmanni type II lectin -- CEL2 Canavalia ensiformis type II lectin -- DGL2 Dioclea grandiflora type II lectin -- CRL2 Cymbosema roseum type II lectin -- DlyL 1Dioclea lasiophylla type I lectin -- DlyL2 Dioclea lasiophylla type II lectin -- GalNAc N-acetyl-d-galactosamine -- HU hemagglutination unit -- MIC minimum inhibitory concentration -- PI unbound material from Sephadex-G50 -- PII unbound material from Sepharose-4B-lactose -- PIII pure lectin -- SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis -- EDTA Ethylenediaminetetraacetic acid -- RP-UPLC reversed-phase chromatography on ultra-performance liquid chromatography -- Q-TOF quadrupole time of flight mass spectrometry -- MS mass spectrometry -- MS/MS tandem mass spectrometry -- PTH Phenylthiohydantoin -- DDA data-dependent acquisition -- CID collision-induced dissociation -- LC50 median lethal concentration -- Gal Galactose -- SEC size exclusion chromatography -- CRD carbohydrate recognition domain -- MBS metal-binding site -- LTA Lotus tetragonolobus lectin -- PNA Arachis hypogaea lectin -- RBL Robinia pseudocacia lectin; Type II lectin -- Dioclea lasiophylla -- Diocleinae; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2020.03.026
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: A novel type II lectin was purified from Dioclea lasiophylla Mart. ex Benth seeds. The activity of DlyL2 was inhibited by N- acetylgalactosamine and α-lactose. DlyL2 is stable in wide range of pH and temperature values. DlyL2 had its primary structure partially characterized. Abstract: A type II lectin, designated as DlyL2, was purified from Dioclea lasiophylla Mart. ex Benth seeds and some of its physicochemical properties determined. The lectin demonstrated specificity for α-lactose and N -acetyl-d -galactosamine and was able to interact with porcine stomach mucin. DlyL2 has 0.78 % carbohydrates in its composition, therefore can be considered a glycoprotein. In addition, its hemagglutinating activity remained stable at a temperature of 90 °C and in a pH range from 5 to 10. Metal chelation treatment did not affect DlyL2 activity suggesting it's not a metalloprotein. Dlyl2 showed an apparent mass of 31 kDa and average molecular mass of 26.371 kDa. Primary structure data could be generated from the partial amino acid sequence of DlyL2, with about 192 residues sequenced by a combination of Edman degradation and tandem mass spectrometry. The lectin is similar to other type II lectins from Diocleinae subtribe, as well as lectins derived from species of more ancient tribes of the Fabaceae family. In addition, DlyL2 exhibited no toxicity to Artemia sp. nauplii. The present study expands the knowledge about the specificity, structural and physicochemical properties of Diocleinae type II lectins.