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Cell Communication and Signaling : CCS Oct 2022Concanavalin A (ConA), the most studied plant lectin, has been known as a potent anti-neoplastic agent for a long time. Since initial reports on its capacity to kill... (Review)
Review
Concanavalin A (ConA), the most studied plant lectin, has been known as a potent anti-neoplastic agent for a long time. Since initial reports on its capacity to kill cancer cells, much attention has been devoted to unveiling the lectin's exact molecular mechanism. It has been revealed that ConA can bind to several receptors on cancerous and normal cells and modulate the related signaling cascades. The most studied host receptor for ConA is MT1-MMP, responsible for most of the lectin's modulations, ranging from activating immune cells to killing tumor cells. In this study, in addition to studying the effect of ConA on signaling and immune cell function, we will focus on the most up-to-date advancements that unraveled the molecular mechanisms by which ConA can induce autophagy and apoptosis in various cancer cell types, where it has been found that P73 and JAK/STAT3 are the leading players. Moreover, we further discuss the main signaling molecules causing liver injury as the most significant side effect of the lectin injection. Altogether, these findings may shed light on the complex signaling pathways controlling the diverse responses created via ConA treatment, thereby modulating these complex networks to create more potent lectin-based cancer therapy. Video Abstract.
Topics: Humans; Concanavalin A; Lectins; Matrix Metalloproteinase 14; Neoplasms; Plant Lectins
PubMed: 36289525
DOI: 10.1186/s12964-022-00972-7 -
Journal of Microbiology and... Dec 2018Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens....
Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens. Structural studies have detailed the interaction between ConA and carbohydrates and the metal coordination environment with manganese and calcium ions (Mn-Ca-ConA). In this study, ConA was crystallized with a cadmium-containing precipitant, and the refined structure indicates that Mn²⁺ was replaced by Cd²⁺ (Cd-Ca-ConA). The structural comparison with ConA demonstrates that the metal-coordinated residues of Cd-Ca-ConA, that is Glu8, Asp10, Asn14, Asp19, and His24, do not have conformational shifts, but residues for sugar binding, including Arg228, Tyr100, and Leu99, reorient their side chains, slightly. Previous studies demonstrated that excess cadmium ions can coordinate with other residues, including Glu87 and Glu183, which were not coordinated with Cd²⁺ in this study. The trimeric ConA in this study coordinated Cd²⁺ with other residues, including Asp80 and Asp82, for the complex generation. The monomers does not have specific interaction near interface regions with the other monomer, but secondary cadmium coordinated with two aspartates (Asp80 and Asp82) from monomer 1 and one aspartate (Asp16) from monomer 2. This study demonstrated that complex generation was induced via coordination with secondary Cd²⁺ and showed the application potential regarding the design of complex formation for specific interactions with target saccharides.
Topics: Binding Sites; Cadmium; Calcium; Canavalia; Concanavalin A; Crystallography, X-Ray; Manganese; Metals; Models, Molecular; Plant Lectins; Protein Binding; Protein Conformation
PubMed: 30380822
DOI: 10.4014/jmb.1809.09027 -
Molecular Systems Biology Apr 2021Sample multiplexing facilitates single-cell sequencing by reducing costs, revealing subtle difference between similar samples, and identifying artifacts such as cell...
Sample multiplexing facilitates single-cell sequencing by reducing costs, revealing subtle difference between similar samples, and identifying artifacts such as cell doublets. However, universal and cost-effective strategies are rather limited. Here, we reported a concanavalin A-based sample barcoding strategy (CASB), which could be followed by both single-cell mRNA and ATAC (assay for transposase-accessible chromatin) sequencing techniques. The method involves minimal sample processing, thereby preserving intact transcriptomic or epigenomic patterns. We demonstrated its high labeling efficiency, high accuracy in assigning cells/nuclei to samples regardless of cell type and genetic background, and high sensitivity in detecting doublets by three applications: 1) CASB followed by scRNA-seq to track the transcriptomic dynamics of a cancer cell line perturbed by multiple drugs, which revealed compound-specific heterogeneous response; 2) CASB together with both snATAC-seq and scRNA-seq to illustrate the IFN-γ-mediated dynamic changes on epigenome and transcriptome profile, which identified the transcription factor underlying heterogeneous IFN-γ response; and 3) combinatorial indexing by CASB, which demonstrated its high scalability.
Topics: Animals; Cell Line, Tumor; Cell Nucleus; Concanavalin A; DNA; DNA Barcoding, Taxonomic; Gene Expression Regulation, Neoplastic; Humans; Mice; RNA-Seq; Single-Cell Analysis; Transcriptome
PubMed: 33821571
DOI: 10.15252/msb.202010060 -
Biomaterials Oct 2018Eye drops producing long-acting ocular anesthesia would be desirable for corneal pain management. Here we present liposome-based formulations to achieve very long ocular...
Eye drops producing long-acting ocular anesthesia would be desirable for corneal pain management. Here we present liposome-based formulations to achieve very long ocular anesthetic effect after a single eye drop instillation. The liposomes were functionalized with succinyl-Concanavalin A (sConA-Lip), which can bind corneal glycan moieties, to significantly prolong the dwell time of liposomes on the cornea. sConA-Lip were loaded with tetrodotoxin and dexmedetomidine (sConA-Lip/TD), and provided sustained release for both. A single topical instillation of sConA-Lip/TD on the cornea could achieve 105 min of complete analgesia and 608 min of partial analgesia, which was significantly longer than analgesia with proparacaine, tetrodotoxin/dexmedetomidine solution or unmodified liposomes containing tetrodotoxin and dexmedetomidine. sConA-Lip/TD were not cytotoxic in vitro to human corneal limbal epithelial cells or corneal keratocytes. Topical administration of sConA-Lip/TD provided prolonged corneal anesthesia without delaying corneal wound healing. Such a formulation may be useful for the management of acute surgical and nonsurgical corneal pain, or for treatment of other ocular surface diseases.
Topics: Administration, Topical; Anesthetics, Local; Animals; Concanavalin A; Cornea; Corneal Keratocytes; Dexmedetomidine; Liposomes; Male; Rats; Rats, Sprague-Dawley; Tetrodotoxin; Wound Healing
PubMed: 30099260
DOI: 10.1016/j.biomaterials.2018.07.054 -
PloS One 2021Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. A... (Comparative Study)
Comparative Study
Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. A recently established method called CUT&Tag uses a Protein-A Tn5 transposase fusion protein, which cuts the genome and inserts adapter sequences nearby the target protein. Throughout most of the CUT&Tag procedure, cells are held on concanavalin A (con A)-conjugated magnetic beads. Proper holding of cells would be decisive for the accessibility of Tn5 to the chromatin, and efficacy of the procedure of washing cells. However, BioMag®Plus ConA magnetic beads, used in the original CUT&Tag protocol, often exhibit poor suspendability and severe aggregation. Here, we compared the BioMag beads and Dynabeads® magnetic particles of which conjugation of con A was done by our hands, and examined the performance of these magnetic beads in CUT&Tag. Among tested, one of the Dynabeads, MyOne-T1, kept excessive suspendability in a buffer even after overnight incubation. Furthermore, the MyOne-T1 beads notably improved the sensitivity in CUT&Tag assay for H3K4me3. In conclusion, the arrangement and the selection of MyOne-T1 refine the suspendability of beads, which improves the association of chromatin with Tn5, which enhances the sensitivity in CUT&Tag assay.
Topics: Animals; Cell Line; Concanavalin A; Epigenomics; HEK293 Cells; Histone Code; Histones; Humans; Immunomagnetic Separation; Magnetic Fields; Methylation; Mice; Particle Size; Recombinant Fusion Proteins; Staphylococcal Protein A; Transposases
PubMed: 34784358
DOI: 10.1371/journal.pone.0259846 -
Analytical Biochemistry Jul 2004Noble metal nanoparticles are well known for their strong interactions with light through the resonant excitations of the collective oscillations of the conduction...
Noble metal nanoparticles are well known for their strong interactions with light through the resonant excitations of the collective oscillations of the conduction electrons on the particles, the so-called surface plasmon resonances. The close proximity of two nanoparticles is known to result in a red-shifted resonance wavelength peak, due to near-field coupling. We have subsequently employed this phenomenon and developed a new approach to glucose sensing, which is based on the aggregation and disassociation of 20-nm gold particles and the changes in plasmon absorption induced by the presence of glucose. High-molecular-weight dextran-coated nanoparticles are aggregated with concanavalin A (Con A), which results in a significant shift and broadening of the gold plasmon absorption. The addition of glucose competitively binds to Con A, reducing gold nanoparticle aggregation and therefore the plasmon absorption when monitored at a near-red arbitrary wavelength. We have optimized our plasmonic-type glucose nanosensors with regard to particle stability, pH effects, the dynamic range for glucose sensing, and the observation wavelength to be compatible with clinical glucose requirements and measurements. In addition, by modifying the amount of dextran or Con A used in nanoparticle fabrication, we can to some extent tune the glucose response range, which means that a single sensing platform could potentially be used to monitor microM --> mM glucose levels in many physiological fluids, such as tears, blood, and urine, where the glucose concentrations are significantly different.
Topics: Biosensing Techniques; Concanavalin A; Dextrans; Glucose; Gold; Nanostructures; Sensitivity and Specificity; Surface Plasmon Resonance
PubMed: 15183773
DOI: 10.1016/j.ab.2004.03.032 -
Pharmaceutical Biology Dec 2022-Propargyl-cysteine (SPRC), an endogenous HS modulator, exerts anti-inflammatory effects on cardiovascular and neurodegenerative disease, but it remains unknown whether...
CONTEXT
-Propargyl-cysteine (SPRC), an endogenous HS modulator, exerts anti-inflammatory effects on cardiovascular and neurodegenerative disease, but it remains unknown whether SPRC can prevent autoimmune hepatitis.
OBJECTIVE
To evaluate the preventive effect of SPRC on concanavalin A (Con A)-induced liver injury and uncover the underlying mechanisms.
MATERIALS AND METHODS
Mice were randomly divided into five groups: control, Con A, SPRC (5 and 10 mg/kg injected intravenously once a day for 7 days), and propargylglycine (PAG; 50 mg/kg injected intraperitoneally 0.5 h before SPRC for 7 days). All mice except the controls were intravenously injected with Con A (20 mg/kg) on day 7. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were evaluated using kits. Inflammatory cytokines (TNF-α and IFN-γ) in the blood and in the liver were detected by ELISA Kit and real-time PCR, respectively. The expression of mitogen-activated protein kinase (MAPK) pathway proteins (p-JNK and p-Akt) and apoptosis proteins (Bax and Bcl-2) was detected using western blotting.
RESULTS
SPRC reduced the levels of AST ( < 0.05) and ALT ( < 0.01) and decreased the release of the inflammatory cytokines. Mechanistically, SPRC increased HS level ( < 0.05) and promoted cystathionine γ-lyase (CSE) expression ( < 0.05). SPRC inhibited the MAPK pathway activation and the apoptosis pathway. All the effects of SPRC were blocked by the CSE inhibitor PAG.
CONCLUSIONS
SPRC prevents Con A-induced liver injury in mice by promoting CSE expression and producing endogenous HS. The mechanisms include reducing the release of inflammatory cytokines, attenuating MAPK pathway activation, and alleviating apoptosis.
Topics: Animals; Chemical and Drug Induced Liver Injury, Chronic; Concanavalin A; Cysteine; Cytokines; Hydrogen Sulfide; Mice; Neurodegenerative Diseases
PubMed: 35701112
DOI: 10.1080/13880209.2022.2080234 -
The Plant Cell Aug 2021Over 30 years ago, an intriguing posttranslational modification was found responsible for creating concanavalin A (conA), a carbohydrate-binding protein from jack bean...
Over 30 years ago, an intriguing posttranslational modification was found responsible for creating concanavalin A (conA), a carbohydrate-binding protein from jack bean (Canavalia ensiformis) seeds and a common carbohydrate chromatography reagent. ConA biosynthesis involves what was then an unprecedented rearrangement in amino-acid sequence, whereby the N-terminal half of the gene-encoded conA precursor (pro-conA) is swapped to become the C-terminal half of conA. Asparaginyl endopeptidase (AEP) was shown to be involved, but its mechanism was not fully elucidated. To understand the structural basis and consequences of circular permutation, we generated recombinant jack bean pro-conA plus jack bean AEP (CeAEP1) and solved crystal structures for each to 2.1 and 2.7 Å, respectively. By reconstituting conA biosynthesis in vitro, we prove CeAEP1 alone can perform both cleavage and cleavage-coupled transpeptidation to form conA. CeAEP1 structural analysis reveals how it is capable of carrying out both reactions. Biophysical assays illustrated that pro-conA is less stable than conA. This observation was explained by fewer intermolecular interactions between subunits in the pro-conA crystal structure and consistent with a difference in the prevalence for tetramerization in solution. These findings elucidate the consequences of circular permutation in the only posttranslation example known to occur in nature.
Topics: Binding Sites; Canavalia; Catalytic Domain; Circular Dichroism; Concanavalin A; Crystallography, X-Ray; Cysteine Endopeptidases; Hydrogen-Ion Concentration; Methylmannosides; Models, Molecular; Protein Conformation; Protein Precursors; Protein Stability; Recombinant Proteins; Solutions
PubMed: 34235541
DOI: 10.1093/plcell/koab130 -
Concanavalin A differentiates gram-positive bacteria through hierarchized nanostructured transducer.Microbiological Research Oct 2021Biosensors are pre-prepared diagnostic devices composed of at least one biological probe. These devices are envisaged for the practical identification of specific...
Biosensors are pre-prepared diagnostic devices composed of at least one biological probe. These devices are envisaged for the practical identification of specific targets of microbiological interest. In recent years, the use of narrow-specific probes such as lectins has been proven to distinguish bacteria and glycoproteins based on their superficial glycomic pattern. For instance, Concanavalin A is a carbohydrate-binding lectin indicated as a narrow-specific biological probe for Gram-negative bacteria. As a drawback, Gram-positive bacteria are frequently overlooked from lectin-based biosensing studies because their identification results in low resolution and overlapped signals. In this work, the authors explore the effect that platform nanostructuration has over the electrochemical response of ConA-based platforms constructed for bacterial detection; one is formed of chitosan-capped magnetic nanoparticles, and another is composed of gold nanoparticle-decorated magnetic nanoparticles. The biosensing platforms were characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) as a function of bacterial concentration. Our results show that probe-target interaction causes variations in the electrical responses of nanostructured transducers. Moreover, the association of gold nanoparticles to magnetic nanoparticles resulted in an electrical enhancement capable of overcoming low resolution and overlapping Gram-positive identification. Both platforms attained a limit of detection of 10 ° CFU mL, which is useful for water analyses and sanitation concerns, where low CFU mL are always expected. Although both platforms were able to detect Gram-negative bacteria, Gram-positives were only correctly differentiated by the gold nanoparticle-decorated magnetic nanoparticles, thus demonstrating the positive influence of hierarchically nanostructured platforms.
Topics: Biosensing Techniques; Concanavalin A; Gold; Gram-Positive Bacteria; Metal Nanoparticles; Transducers
PubMed: 34364021
DOI: 10.1016/j.micres.2021.126834 -
Mediators of Inflammation 20224-Octyl itaconate (OI) is a novel anti-inflammatory metabolite that exerts protective effects in many various disease models. However, its function in autoimmune...
4-Octyl itaconate (OI) is a novel anti-inflammatory metabolite that exerts protective effects in many various disease models. However, its function in autoimmune hepatitis- (AIH-) associated hepatic injury has not been investigated. In this study, we successfully used concanavalin A (Con A) to establish an AIH-associated liver injury model. Furthermore, we investigated the effect of OI in Con A-induced liver injury and found that OI mitigated Con A-induced histopathological damage. OI administration reduced serum levels of alanine transaminase and aspartate transaminase in Con A-treated mice and attenuated the infiltration of macrophages induced by Con A. Moreover, OI effectively inhibited the expression of proinflammatory cytokines including interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-), interferon-gamma (IFN-), and IL-1 induced by Con A. Furthermore, OI decreased hepatocyte apoptosis and malondialdehyde levels and increased the reduced glutathione/oxidized glutathione ratio in the Con A-induced liver injury model. In addition, we found that OI inhibited Con A-induced hepatocyte apoptosis in vitro, while Nrf2 deletion eliminated this effect. Furthermore, we administrated the Nrf2 inhibitor ML385 in OI+Con A-treated mice and found that ML385 eliminated the protective effect of OI in vivo. In addition, OI inhibited Con A-induced activation of nuclear factor-kappa B (NF-𝜅B) and the expression of proinflammatory cytokines in macrophages. Therefore, OI protected mice from Con A-induced liver damage and may be associated with Nrf2 activation and NF-𝜅B inhibition. Finally, our study revealed that OI inhibited TNF-, or supernatants from Con A-treated RAW264.7 cells induced hepatocyte apoptosis. In conclusion, our study indicated that OI alleviated Con A-induced hepatic damage by reducing inflammatory response, oxidative stress, and apoptosis.
Topics: Animals; Concanavalin A; Hepatitis, Autoimmune; Mice; Succinates
PubMed: 35310452
DOI: 10.1155/2022/5766434