Lectins are glycan-binding proteins that play a very important role in biomolecule recognition, cell communication, immune regulation, and protein transport. Lectins have been characterized in legumes, Solanaceae, etc. CD BioGlyco provides clients with professional Lectin Production Services of Natural Origin such as plant origin, animal origin, and fungal origin. D. stramonium, also known as thorn apple and jimson weed, is an annual tropical herb of the genus Datura in the Solanaceae family. DSL is extracted and purified from this plant of the Solanaceae family, specifically binds chitin, and has antimicrobial and anti-tumor activity. The carbohydrate-binding site of this lectin preferentially recognizes complex N-glycans containing three or more branches of N-acetyllactosamine moieties rather than single N-acetylglucosamine residues. Among three-branch complex N-glycans, DSL preferentially binds to C-2,6-branched rather than C-2,4-branched N-glycans. Studies have found that β1-4-linked N-acetyllactosamine or N-acetylglucosamine oligomers inhibit hemagglutination activity.
Fig.1 DSL production service. (CD BioGlyco)
D. stramonium seed is ground in a seed mill and extracted with phosphate-buffered saline. The obtained crude cell-free extract is centrifuged, the supernatant is taken, and the precipitate is collected after treatment with acetone. The precipitate is resuspended in phosphate buffer, dialyzed, and purified by affinity chromatography.
Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is performed using a precast homogeneous gel, and proteins in the gel are stained with Coomassie brilliant blue (CBB). The molecular weight of DSL is estimated at the constructed calibration curve of the marker.
To determine the N-terminal sequence of DSL subunits, each subunit band electroblotted onto a polyvinylidene difluoride (PVDF) membrane is excised and analyzed with a protein sequencer.
Agglutination is one of the easiest immunological tests to perform and is used to detect antigens or antibodies. We test for hemagglutination by serially diluting the sample to be tested in phosphate buffer and then mixing it with an equal volume of rabbit red blood cell suspension.
We also provide clients with a variety of DSL products to choose from, including but not limited to:
Technology: SDS-PAGE, Amino Acid Sequence
Journal: Glycobiology
IF: 4.3
Published: 2015
Results: DSL is composed of three isolectins, either as homo- or heterodimers of the A and B subunits. To obtain the structural information required for molecular cloning, the authors purified three D. stramonium isolectin fractions from Jimson weed and subjected the fractions to SDS-PAGE, amino acid sequencing, and cDNA cloning. The results showed that part of the amino acid sequence of purified DSL-B was identical to part of the amino acid sequence deduced from the cDNA sequence, that is, the cloned cDNA encoded DSL-B.
Fig.2 SDS-PAGE and nucleotide sequence of DSL. (Nishimoto, et al., 2015)
CD BioGlyco provides a broad range of Lectin Production Services to identify specific carbohydrates to support your glycomics and glycoproteomics research. Please feel free to contact us if you have any lectin production needs.
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