Lectins purified from medicinal and edible mushrooms: Insights into their antiviral activity against pathogenic viruses.

For thousands of years, fungi have been a valuable and promising source of therapeutic agents for treatment of various diseases. Mushroom is a macrofungus which has been cultivated worldwide for its nutritional value and medicinal applications. Several bioactive molecules were extracted from mushroom such as polysaccharides, lectins and terpenoids. Lectins are carbohydrate-binding proteins with non-immunologic origin. Lectins were classified according to their structure, origin and sugar specificity. This protein has different binding specificity with surface glycan moiety which determines its activity and therapeutic applications. A wide range of medicinal activities such as antitumor, antiviral, antimicrobial, immunomodulatory and antidiabetic were reported from sugar-binding proteins. However, glycan-binding protein from mushroom is not well explored as antiviral agent. The discovery of novel antiviral agents is a public health emergency to overcome the current pandemic and be ready for the upcoming viral pandemics. The mechanism of action of lectin against viruses targets numerous steps in viral life cycle such as viral attachment, entry and replication. This review described the history, classification, purification techniques, structure-function relationship and different therapeutic applications of mushroom lectin. In addition, we focus on the antiviral activity, purification and physicochemical characteristics of some mushroom lectins.

LectomeXplore, an update of UniLectin for the discovery of carbohydrate-binding proteins based on a new lectin classification.

Lectins are non-covalent glycan-binding proteins mediating cellular interactions but their annotation in newly sequenced organisms is lacking. The limited size of functional domains and the low level of sequence similarity challenge usual bioinformatics tools. The identification of lectin domains in proteomes requires the manual curation of sequence alignments based on structural folds. A new lectin classification is proposed. It is built on three levels: (i) 35 lectin domain folds, (ii) 109 classes of lectins sharing at least 20% sequence similarity and (iii) 350 families of lectins sharing at least 70% sequence similarity. This information is compiled in the UniLectin platform that includes the previously described UniLectin3D database of curated lectin 3D structures. Since its first release, UniLectin3D has been updated with 485 additional 3D structures. The database is now complemented by two additional modules: PropLec containing predicted ß-propeller lectins and LectomeXplore including predicted lectins from sequences of the NBCI-nr and UniProt for every curated lectin class. UniLectin is accessible at https://www.unilectin.eu/.

Structural Database for Lectins and the UniLectin Web Platform.

The search for new biomolecules requires a clear understanding of biosynthesis and degradation pathways. This view applies to most metabolites as well as other molecule types such as glycans whose repertoire is still poorly characterized. Lectins are proteins that recognize specifically and interact noncovalently with glycans. This particular class of proteins is considered as playing a major role in biology. Glycan-binding is based on multivalence, which gives lectins a unique capacity to interact with surface glycans and significantly contribute to cell-cell recognition and interactions. Lectins have been studied for many years using multiple technologies and part of the resulting information is available online in databases. Unfortunately, the connectivity of these databases with the most popular omics databases (genomics, proteomics, and glycomics) remains limited. Moreover, lectin diversity is extended and requires setting out a flexible classification that remains compatible with new sequences and 3D structures that are continuously released. We have designed UniLectin as a new insight into the knowledge of lectins, their classification, and their biological role. This platform encompasses UniLectin3D, a curated database of lectin 3D structures that follows a periodically updated classification, a set of comparative and visualizing tools and gradually released modules dedicated to specific lectins predicted in sequence databases. The second module is PropLec, focused on ß-propeller lectin prediction in all species based on five distinct family profiles. This chapter describes how UniLectin can be used to explore the diversity of lectins, their 3D structures, and associated functional information as well as to perform reliable predictions of ß-propeller lectins.

Expression of Lectins in Heterologous Systems.

Lectins are proteins that have the ability to recognize and bind in a reversible and specific way to free carbohydrates or glycoconjugates of cell membranes. For these reasons, they have been extensively used in a wide range of industrial and pharmacological applications. Currently, there is great interest in their production on a large scale. Unfortunately, conventional techniques do not provide the appropriate platform for this purpose and therefore, the heterologous production of lectins in different organisms has become the preferred method in many cases. Such systems have the advantage of providing better yields as well as more homogeneous and better-defined properties for the resultant products. However, an inappropriate choice of the expression system can cause important structural alterations that have repercussions on their biological activity since the specificity may lay in their post-translational processing, which depends largely on the producing organism. The present review aims to examine the most representative studies in the area, exposing the four most frequently used systems (bacteria, yeasts, plants and animal cells), with the intention of providing the necessary information to determine the strategy to follow in each case as well as their respective advantages and disadvantages.

Glycan diversity in the vomeronasal organ of the Korean roe deer, Capreolus pygargus: A lectin histochemical study.

Glycans in the epithelium play an important role in cell-to-cell communication and adhesion. No detailed evaluation of glycoconjugates in the vomeronasal organs (VNO) of the roe deer has been published previously. The aim of this study was to characterize glycan epitopes in the vomeronasal sensory epithelium (VSE) and non-sensory epithelium (VNSE) using lectin histochemistry. Glycan epitopes identified by lectin histochemistry were grouped as follows: N-acetylglucosamine (s-WGA, WGA, BSL-II, DSL, LEL, STL), mannose (Con A, LCA, PSA), galactose (RCA120, BSL-I, Jacalin, PNA, ECL), N-acetylgalactosamine (VVA, DBA, SBA, and SJA), fucose (UEA-I) and complex type N-glycan (PHA-E and PHA-L) groups. The free border of the VSE was positive for all 21 lectins, and 18 of the lectins (excluding DBA, SJA, and PHA-L) showed weak and/or moderate staining in the receptor cells. The supporting cells were weakly positive for 19 lectins (excluding PNA and SJA). Moreover, 17 lectins (excluding BSL-II, Jacalin, PNA, and SJA) were expressed in the basal cells. In the VNSE of roe deer, the free border showed staining for all 21 lectins examined. The ciliated cells were positive for 16 lectins (excluding BSL-II, DSL, PNA, VVA, and SJA). Furthermore, 15 lectins (excluding DSL, LEL, ECL, UEA-I, PHA-E, and PHA-L) were expressed in goblet cells. Twenty lectins (excluding SJA) were expressed in the acini of the vomeronasal glands. Collectively, both VSE and VNSE were rich in N-acetylglucosamine, mannose, galactose, N-acetylgalactosamine, fucose, and complex-type N-glycans, although the different cell types of the VSE and VNSE expressed different glycoconjugates of varying intensities, suggesting that these carbohydrate residues may be involved in odor perception as well as cell-to-cell communication in the VNO.

A novel immune-tolerable and permeable lectin-like protein from mushroom Agaricus bisporus.

A lectin like protein designated as LSMT is recently discovered in Agaricus bisporus. The protein adopts very similar structure to Ricin-B like lectin from Clitocybe nebularis (CNL) and HA-33 from Clostridium botulinum (HA-33), which both recognize sugar molecules that decorate the surface of the epithelial cells of the intestine. A preliminary study in silico pointed out potential capability of LSMT to perform such biological activity. Following that hypothesis, we demonstrated that LSMT is indeed capable of penetrating out from a dialysis tube of the mice intestine origin. Furthermore, the protein appeared not to evoke the immune response upon introduction into mice, unlike its structural homologs. This is the first report on the biological implication of LSMT that might lead to its application.

Hemagglutinating/Hemolytic activities in extracts of marine invertebrates from the Brazilian coast and isolation of two lectins from the marine sponge Cliona varians and the sea cucumber Holothuria grisea.

Twenty species of marine invertebrates from the Brazilian coast were screened for hemagglutinating/hemolytic activity. In at least twelve tested species, hemagglutinating activity was different for different blood types, suggesting the presence of lectins. Extracts from four species showed hemolytic activity. Two new lectins were purified from the marine sponge Cliona varians (CvL-2) and sea cucumber Holothuria grisea (HGL). CvL-2 was able to agglutinate rabbit erythrocytes and was inhibited by galactosides. The hemagglutinating activity was optimal in pH neutral and temperatures below 70 °C. CvL-2 is a trimeric protein with subunits of 175 kDa. On the other hand, HGL showed both hemagglutinating and hemolytic activity in human and rabbit erythrocytes, but hemolysis could be inhibited by osmotic protection, and agglutination was inhibited by mucin. HGL was stable in pH values ranging from 4 to 10 and temperatures up to 90 °C. In electrophoresis and gel filtration, HGL was a monomeric protein with 15 kDa. CvL-2 and HGL showed different levels of toxicity to Artemia naplii. CvL-2 showed LC50 of 850.1 µg/mL, whereas HGL showed LC50 of 9.5 µg/mL.

The Lectin Frontier Database (LfDB), and data generation based on frontal affinity chromatography.

Lectins are a large group of carbohydrate-binding proteins, having been shown to comprise at least 48 protein scaffolds or protein family entries. They occur ubiquitously in living organisms-from humans to microorganisms, including viruses-and while their functions are yet to be fully elucidated, their main underlying actions are thought to mediate cell-cell and cell-glycoconjugate interactions, which play important roles in an extensive range of biological processes. The basic feature of each lectin's function resides in its specific sugar-binding properties. In this regard, it is beneficial for researchers to have access to fundamental information about the detailed oligosaccharide specificities of diverse lectins. In this review, the authors describe a publicly available lectin database named "Lectin frontier DataBase (LfDB)", which undertakes the continuous publication and updating of comprehensive data for lectin-standard oligosaccharide interactions in terms of dissociation constants (Kd's). For Kd determination, an advanced system of frontal affinity chromatography (FAC) is used, with which quantitative datasets of interactions between immobilized lectins and >100 fluorescently labeled standard glycans have been generated. The FAC system is unique in its clear principle, simple procedure and high sensitivity, with an increasing number (>67) of associated publications that attest to its reliability. Thus, LfDB, is expected to play an essential role in lectin research, not only in basic but also in applied fields of glycoscience.

Purification of a secreted lectin from Andrias davidianus skin and its antibacterial activity.

A lectin secreted from Andrias davidianus skin (ADL) was purified by affinity chromatography on porcine stomach mucin (type III) (PSM)-crosslinked albumin, followed by gel filtration on Sephadex G-100 and HPLC on TSK gel G3000PWXL. The purified lectin was found to be a dimeric protein, as revealed by SDS-PAGE and MALDI-TOF analysis. SDS-PAGE showed that the ADL protein had a molecular mass of 17 kDa. ADL produced an 8.5 kDa band when examined using SDS-PAGE under reducing conditions. ADL agglutinated native and trypsinized human B erythrocytes. The hemagglutination activity was inhibited by glycoproteins, such as PSM and asialo-PSM, but not by any of the monosaccharides tested. The activity was stable between 4 °C and 50 °C. Significant ADL activity was observed between pH 4­5. The lectin reaction did not depend on the presence of the divalent cation Ca2+ or Mg2+. The N-terminal ADL sequence was determined to be VGYTVGATPM. The lectin exhibited antibacterial activity, involving growth and respiration inhibition in Escherichia coli, Enterobacter aerogenes, Staphylococcus aureus, Bacillus subtilis and Shewanella sp. Furthermore, ADL showed inhibition activity against the yeast Saccharomyces cerevisiae. These findings suggest that ADL plays an important role in the innate immunity of A. davidianus on the body surface.

Lectin-like molecules in transcriptome of Littorina littorea hemocytes.

The common periwinkle Littorina littorea was introduced in the list of models for comparative immunobiology as a representative of phylogenetically important taxon Caenogastropoda. Using Illumina sequencing technology, we de novo assembled the transcriptome of Littorina littorea hemocytes from 182 million mRNA-Seq pair-end 100 bp reads into a total of 15,526 contigs clustered in 4472 unigenes. The transcriptome profile was analyzed for presence of carbohydrate-binding molecules in a variety of architectural contexts. Hemocytes' repertoire of lectin-like proteins bearing conserved carbohydrate-recognition domains (CRDs) is highly diversified, including 11 of 15 lectin families earlier described in animals, as well as the novel members of lectin family found for the first time in mollusc species. The new molluscan lineage-specific domain combinations were confirmed by cloning and sequencing, including the fuco-lectin related molecules (FLReMs) composed of N-terminal region with no sequence homology to any known protein, a middle Fucolectin Tachylectin-4 Pentaxrin (FTP) domain, and a C-terminal epidermal growth factor (EGF) repeat region. The repertoire of lectin-like molecules is discussed in terms of their potential participation in the receptor phase of immune response. In total, immune-associated functions may be attributed to 70 transcripts belonging to 6 lectin families. These lectin-like genes show low overlap between species of invertebrates, suggesting relatively rapid evolution of immune-associated genes in the group. The repertoire provides valuable candidates for further characterization of the gene functions in mollusc immunity.

Lectin structures: classification based on the 3-D structures.

Recent progress in structural biology has elucidated the three-dimensional structures and carbohydrate-binding mechanisms of most lectin families. Lectins are classified into 48 families based on their three-dimensional structures. A ribbon drawing gallery of the crystal and solution structures of representative lectins or lectin-like proteins is appended and may help to convey the diversity of lectin families, the similarity and differences between lectin families, as well as the carbohydrate-binding architectures of lectins.

Lectins in human pathogenic fungi / Lectinas en hongos patógenos para el ser humano

Lectins are carbohydrate-binding proteins widely distributed in nature. They constitute a highly diverse group of proteins consisting of many different protein families that are, in general, structurally unrelated. In the last few years, mushroom and other fungal lectins have attracted wide attention due to their antitumour, antiproliferative and immunomodulatory activities. The present mini-review provides concise information about recent developments in understanding lectins from human pathogenic fungi. A bibliographic search was performed in the Science Direct and PubMed databases, using the following keywords "lectin", "fungi", "human" and "pathogenic". Lectins present in fungi have been classified; however, the role played by lectins derived from human pathogenic fungi in infectious processes remains uncertain; thus, this is a scientific field requiring more research. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012) (AU)

Las lectinas son proteínas que se unen a los hidratos de carbono y están ampliamente distribuidas en la naturaleza. Constituyen un grupo muy diverso de proteínas incluidas en muchas familias que en general carecen de relación estructural. En los últimos años, se ha prestado mucha atención a las lectinas fúngicas debido a sus actividades antitumorales, antiproliferativas e inmunomoduladoras. La presente revisión proporciona información sucinta sobre los progresos recientes acontecidos en la comprensión de estas moléculas a partir de hongos patógenos para el ser humano. Emprendimos una búsqueda bibliográfica de los estudios publicados en las bases de datos Science Direct y PubMed, usando las palabras claves: «lectin» (lectina), «fungi» (hongos), «human» (humano) y «pathogenic» (patogénico). Se han clasificado las lectinas presentes en los hongos; sin embargo, el papel que desempeñan en los procesos infecciosos de hongos patógenos para el ser humano sigue por dilucidar, por lo que este es un ámbito científico que requiere mayor investigación.Este manuscrito forma parte de la serie de artículos presentados en el «V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi» (Oaxaca, México, 2012) (AU)

Lectins in human pathogenic fungi.

Lectins are carbohydrate-binding proteins widely distributed in nature. They constitute a highly diverse group of proteins consisting of many different protein families that are, in general, structurally unrelated. In the last few years, mushroom and other fungal lectins have attracted wide attention due to their antitumour, antiproliferative and immunomodulatory activities. The present mini-review provides concise information about recent developments in understanding lectins from human pathogenic fungi. A bibliographic search was performed in the Science Direct and PubMed databases, using the following keywords "lectin", "fungi", "human" and "pathogenic". Lectins present in fungi have been classified; however, the role played by lectins derived from human pathogenic fungi in infectious processes remains uncertain; thus, this is a scientific field requiring more research. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012).

Classification of lectins by pattern recognition using glyconanoparticles.

Carbohydrate-functionalized gold nanoparticles were employed to differentiate plant-legume lectins using a statistical analysis method of linear discriminant analysis (LDA). Various carbohydrates were conjugated on gold nanoparticles, and the resulting glyconanoparticles were treated with lectins. Changes in the localized surface plasmon resonance of the glyconanoparticles upon lectin binding were recorded, and the data were subjected to LDA. Results showed that the glyconanoparticles successfully differentiated all lectins.

Fish lily type lectin-1 contains ß-prism architecture: immunological characterization.

In this study we report a full-length lily type lectin-1 (CsLTL-1) identified from striped murrel, Channa striatus. CsLTL-1 was identified from the established C. striatus cDNA library using GS-FLX™ genome sequencing technology and was found to contain 354 nucleotide base pairs and its open reading frame (ORF) encodes a 118 amino acid residue. CsLTL-1 mRNA is predominately expressed in the gills and is up-regulated upon infection with fungus (Aphanomyces invadans) and bacteria (Aeromonas hydrophila). Hemagglutination studies with recombinant CsLTL-1 show that, at 4µg/ml agglutinates occurs in a calcium independent manner and is inhibited in the presence of d-mannose (50mM) and d-glucose (100mM). The CsLTL-1 sequence was completely characterized using various bioinformatics tools. CsLTL-1 peptide contains a mannose binding site at 30-99 along with its specific motif of ß-prism architecture. The phylogenetic analysis showed that CsLTL-1 clustered together with LTL-1 from Oplegnathus fasciatus. CsLTL-1 protein 3D structure was predicted by I-Tasser program and the model was evaluated using Ramachanran plot analysis. The secondary structure analysis of CsLTL-1 reveals that the protein contains 23% ß-sheets and 77% coils. The overall results showed that CsLTL-1 is an important immune gene involved in the recognition and elimination of pathogens in murrels.

Comparative genomic and phylogenetic analyses of the intelectin gene family: implications for their origin and evolution.

Intelectin is a newly characterized gene family involved in early embryogenesis, host-pathogen interactions and iron metabolism. In this study, we searched the genomes of metazoans by extensive BLAST survey and found no intelectin homologs in invertebrate metazoans but 12 in amphioxus Branchiostoma floridae and 21 in ascidians Ciona intestinalis. Some ascidians oocyte cortical granule lectins (CGLs) have unknown insertion sequences between fibrinogen-related domain (FReD) and Intelectin Domain, the boundaries of which are equivalent to exon structures. In addition to ascidians intelectins/CGLs located in the base, phylogenetic tree comprises four main clades representing mammal, frog, fish, and amphioxus, indicating that intelectin genes undergo extensive lineage-specific duplication or gene conversion. However, genomic neighborhood surrounding analysis shows that clear proto-orthologies are difficult to be established among these counterparts. In addition, sequence comparison and phylogenetic analysis of FReDs from intelectins and other fibrinogen-like proteins from choanoflagellate, anemone, frog and human indicate FReDs of intelectins are unique. Likewise, these choanoflagellate and anemone genes may be close to intelectin gene.

Siglecs and immune regulation.

Sialic acid-binding Ig-like lectins, or Siglecs, vary in their specificity for sialic acid-containing ligands and are mainly expressed by cells of the immune system. Many Siglecs are inhibitory receptors expressed in innate immune cells that regulate inflammation mediated by damage-associated and pathogen-associated molecular patterns (DAMPs and PAMPs). This family also includes molecules involved in adhesion and phagocytosis and receptors that can associate with the ITAM-containing DAP12 adaptor. Siglecs contribute to the inhibition of immune cells both by binding to cis ligands (expressed in the same cells) and by responding to pathogen-derived sialoglycoconjugates. They can help maintain tolerance in B lymphocytes, modulate the activation of conventional and plasmacytoid dendritic cells, and contribute to the regulation of T cell function both directly and indirectly. Siglecs modulate immune responses, influencing almost every cell in the immune system, and are of relevance both in health and disease.

Structural and functional diversity of the lectin repertoire in teleost fish: relevance to innate and adaptive immunity.

Protein-carbohydrate interactions mediated by lectins have been recognized as key components of innate immunity in vertebrates and invertebrates, not only for recognition of potential pathogens, but also for participating in downstream effector functions, such as their agglutination, immobilization, and complement-mediated opsonization and killing. More recently, lectins have been identified as critical regulators of mammalian adaptive immune responses. Fish are endowed with virtually all components of the mammalian adaptive immunity, and are equipped with a complex lectin repertoire. In this review, we discuss evidence suggesting that: (a) lectin repertoires in teleost fish are highly diversified, and include not only representatives of the lectin families described in mammals, but also members of lectin families described for the first time in fish species; (b) the tissue-specific expression and localization of the diverse lectin repertoires and their molecular partners is consistent with their distinct biological roles in innate and adaptive immunity; (c) although some lectins may bind endogenous ligands, others bind sugars on the surface of potential pathogens; (d) in addition to pathogen recognition and opsonization, some lectins display additional effector roles, such as complement activation and regulation of immune functions; (e) some lectins that recognize exogenous ligands mediate processes unrelated to immunity: they may act as anti-freeze proteins or prevent polyspermia during fertilization.

Lectins of beneficial microbes: system organisation, functioning and functional superfamily.

In this review our last results and proposals with respect to general aspects of lectin studies are summarised and compared. System presence, organisation and functioning of lectins are proposed, and accents on beneficial symbiotic microbial lectins studies are presented. The proposed general principles of lectin functioning allows for a comparison of lectins with other carbohydrate-recognition systems. A new structure-functional superfamily of symbiotic microbial lectins is proposed and its main properties are described. The proposed superfamily allows for extended searches of the biological activities of any microbial member. Prospects of lectins of beneficial symbiotic microorganisms are discussed.

Lectins of living organisms. The overview.

Occurrence, organization, and functioning of lectins as well as their current classifications are under investigation. Results indicate importance of symbiotic lectins for clinical microecology. Lectins and lectin-based approaches have wide perspectives for medical biotechnology. Lectin terms, relationships between lectins and enzymes are discussed.