Dioclea Grandiflora Lectin Production Service
Dioclea grandiflora Lectin (DGL) Production Service at CD BioGlyco
The specific carbohydrate recognition properties of lectins make them important tools in glycomic and glycoproteomic research. It selectively and specifically recognizes different carbohydrates without changing the covalent structure of glycosyl ligands, and plays an important role in protein-cell and cell-cell interactions. For many years, CD BioGlyco has been focusing on glycobiology research and has accumulated rich experience and expertise in the analysis, synthesis, extraction, separation, purification, and characterization of carbohydrates. Our products and services are for research use only and are not intended for therapeutic use.
Concanavalin A (ConA) is a well-known carbohydrate-binding protein derived from leguminous plants with a high affinity for terminal mannose (Man) or glucose (Glc) residues. DGL is a lectin with very similar physical properties and structural characteristics to ConA, isolated from the seeds of the leguminous plant D. grandiflora. The two lectins share 81% amino acid sequence identity, and the amino acid residues of the sugar-binding site are conserved in both ConA and DGL. CD BioGlyco provides clients with extraction, purification, characterization, bioactivity testing, and conjugation services of Plant-derived Lectins ranging from milligrams to grams. The highly purified DGL we produce is used to study specific glycans and glycosylation patterns in glycoproteins.
Fig.1 DGL production service. (CD BioGlyco)
- Purification of DGL
Crude DGL is continuously extracted from D. grandiflora seeds using specific reagents, and different fractions are obtained by Affinity Chromatography, dialyzed, and freeze-dried to obtain DGL powder.
- General Determination of DGL
- An amino acid analyzer is used to determine the amino acid composition of purified DGL.
- Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is used to determine the molecular mass of lectins.
- Use D-Glc as the standard to determine the neutral sugar content in DGL.
- The metal content of lectins is determined using an atomic absorption spectrophotometer.
- The glycoprotein properties of DGL are evaluated using periodate-Schiff and ConA-peroxidase assessments.
- Minimum inhibitory concentration determinations are performed using lactose, melezitose, and sucrose as well as pure DGL and A+ red blood cells.
- Custom Lectin Conjugates
Fluorescently labeled lectins are used to localize and detect specific carbohydrates on the cell surface or within cells, making them versatile reagents in microscopy imaging. CD BioGlyco uses special conjugation procedures to provide clients with DGL conjugation services with a variety of fluorescent dyes, such as tetramethylrhodamine (TRITC), fluorescein (FITC), Texas Red, Alexa Fluor, DyLight 488, etc. We also provide custom colloidal gold conjugation services with particle sizes ranging from 5 to 40 nm.
In addition, we also provide clients with a variety of DGL products to choose from, including but not limited to:
Publication
Technology: X-ray diffraction
Journal: FEBS Letters
IF: 3.5
Published: 2015
Results: The authors studied the pH dependence and crystal structure of recombinant wild-type and site-directed mutants of the pH-stable tetrameric DGL by equilibrium sedimentation. The results indicated that histidine residues 51 and 131 played an important role in the pH-dependent dimer-tetramer transition. H131 affected the network of loop 114-125 residues of all four subunits in the central cavity, and H51 formed H131-mediated dimer contacts through the central cavity, thereby stabilizing the structure of the tetramer.
Fig.2 Equilibrium sedimentation analysis and crystal structure. (Zamora-Caballero, et al., 2015)
Advantages
- Use affinity chromatography, Gel Chromatography, and other techniques to purify different fractions of crude lectin.
- Help clients analyze the molecular mass, neutral sugar content, metal content, glycoprotein characteristics, amino acid composition, and minimum inhibitory concentration of purified DGL.
- We not only label DGL with biotin, fluorescent dyes, etc. but also provide colloidal gold-labeled DGL conjugates with high quality and precise optical density.
- We use a special conjugation procedure to optimize the binding of DGL to dye molecules and remove unbound fluorescent dyes and uncoupled lectins while maintaining a high signal-to-noise ratio.
CD BioGlyco has many years of experience and expertise in Lectin Production, providing clients with unconjugated, biotin-conjugated, fluorescein-conjugated, and colloidal gold-labeled lectins from milligram to gram quantities. If you have any lectin needs in glycobiology research, please feel free to contact us and we will be your best partner.
Reference
- Zamora-Caballero, S.; et al. Quaternary structure of Dioclea grandiflora lectin assessed by equilibrium sedimentation and crystallographic analysis of recombinant mutants. FEBS Letters. 2015, 589(18): 2290-2296.
