The binding of lectins to carbohydrate moieties on haemocytes of the insects, Blaberus craniifer (Dictyoptera) and Extatosoma tiaratum (Phasmida) (original) (raw)
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Insect Biochemistry and Molecular Biology, 1998
A lectin specific for -1,3-glucans has previously been purified from the cockroach, Blaberus discoidalis. Using polyclonal antiserum against the lectin, similar molecules with agglutinating activity were detected in the sera of several cockroach species, namely, B. craniifer, Leucophaea maderae, Gromphadorhina portentosa, and Periplaneta americana, by various methods including immunodiffusion, immunoelectrophoresis and Western blotting. Immunoaffinity chromatography using purified antibody against the B. discoidalis -1,3-glucan-specific lectin immobilized on CNBr-activated Sepharose 4B matrix was successfully employed to purify the equivalent molecules from sera of these cockroaches. The purified lectins have molecular masses of ca. 520 kDa by gel filtration and subunit mass estimates of 80-82 kDa by SDS-PAGE. Moreover, they have a similar overall structure to B. discoidalis lectin when examined by electron microscopy. The purified lectin enhanced the activation of prophenoloxidase by laminarin. Localization of the -1,3-glucan-specific lectin using antibody against the B. discoidalis lectin showed that the molecule was associated with the haemocyte surface as well as in the cytoplasm of these cells of B. craniifer, L. maderae and P. americana.
Developmental & Comparative Immunology, 1989
OAbstract-CeJlular lectins (CLs) of Phallusin mamillata were demonstrated in protein prepa rations obtained by salt fractionation from he mocytes sonicated in a suitable medium. Since the lectins from the precipitated fraction bind sugars containing D-galactosyl groups, they were purified by affmity chromatography on Sepharose. SDS-PAGE under reducing condi tions showed that CLs are formed of two com ponents of apparent MWs approximately 36,900 and 35,090 and thus differ from serum lectins (SLs) (MW about 62,2(}O). The "shrinkage" observed when SLs were exam ined under nonreducing conditions suggest the presence of intrachain disulpbide bonds which can affect the molecular structure of the SLs. CL-SL differences were also revealed by the nonidentity reaction of the immuno-precipitate in immunodiffusion using 'an anti-SL immune serum. The capacity of hemocytes~oform.~ found on hemocyte surfaces in some species (11,13,14,15). The plasma of tunicates contains lectins characterized by binding of specific sites to substances such as sialic acid and, more frequently, D-galactosyl residues. Lectins have been detected by their capacity to aggJu tinate untreated and enzyme-treated vertebrate erythrocytes and so far their biological role has not been determined. We recently reported that D-galactose specific lectins of Ascidia malaca can be found both free in the serum and present on hemocyte surface (4,16). This finding suggests that lee tins on cell surfaces might act as receptors which can bind di rectly with surface sugars on foreign particles or identify self-markers on cell settes or clumps with erytbrocytes "dem~i:t->~embranes.. .. . strated that they possess Cl-Iactose specific 'Ct.s;.. ><~feJIular recognItIon In Invertebrates on their surfaces. OKeywords-Tunicates; Hemocytes; Lectins.
Journal of Asia-Pacific Entomology
Lectins due to their affinity to carbohydrate moiety are involved in diverse functions like cell attachment in embryogenesis, organogenesis and cellular trafficking as well as nonself recognition in immune responses. Agglutinating activity was detectable in Plutella xylostella (Yponomeutidae: Lepidoptera) against 14 different species including bacterial and yeast cells, among which the whole body homogenate of P. xylostella agglutinated Providencia vermicola, Flavobacterium sp., and Saccharomyces cerevisiae with high titers. On analysis of physico-chemical properties, this putative agglutinating factor (s) was specifically dependent on the presence of Ca++ for its activity and was reversibly sensitive to EDTA. The agglutinating activity was stable at pH 6–8, but was heat-labile. The agglutinating factor (s) was proteinaceous in nature as it was completely precipitable by ammonium sulphate. Its carbohydrate binding activity was demonstrated by inhibition assay, which revealed that me...
Lectins with Insecticidal and Insectistatic Activities
2018
Lectins are an important group of proteins which are spread in all kingdoms of life. Their most lighted characteristic is associated to their specific carbohydrate binding, although function has been not even identified. According to their carbohydrate specificity, several biological activities have been assessed, finding that lectins can be used as mitogenic agents, biomarkers, and cytotoxic and insecticide proteins. Lectins have been classified according to several features such as structure, source, and carbohydrate recognition. The Protein Research Group (PRG) has worked on Colombian seeds from the family of Fabaceae and Lamiaceae plants, isolating and characterizing their lectins, and found more than one lectin in some plants, indicating that according to its specificity, different lectins can have different biological activities. In the case of legume domain lectins, they have shown the biggest potential as insecticide or insectistatic agents due to the glycosylation pattern i...
Lectin Histochemistry of Agrotis segetum Midgut Cells and Peritrophic Membrane
2018
Midgut cell and peritrophic membranes of grain pest Agrotis segetum larvae were examined by light microscope. Six biotinylated lectins were used in the assays by applying histochemical methods (Avidin-Biotin-Peroxidase). The aim of this study was to find lectins binding on peritrophic membranes if they were available as insecticidal agents. Results of the study indicated that lectins BPA ( Bauhinia purpurea ) and GS-I ( Griffonia simplicifolia ) strongly stained whereas, PNA (Pea Nut Agglutinin) and UEA-1 ( Ulex europaeus ) moderately stained the tissues. However, WGA (Wheat Germ Agglutinin) and Con-A ( Canavalia ensiformis ) stained the tissues weakly. The common feature of two binding proteins, BPA and GS-I lectin was binding to D- galactose. Our examinations revealed that D-galactose mostly exists in A. segetum larvae midgut cell membranes and peritrophic membrane and this means BPA and GS-I lectins can be used as insecticidal agents against A. segetum .
Biochemical Journal, 1993
Three agglutinins (lectins), designated BDLl, BDL2 and BDL3, were identified in the haemolymph of the cockroach Blaberus discoidalis by erythrocyte cross-adsorption and sugar inhibition tests. With the use of (NH4)2SO4 fractionation, anion-exchange and affinity chromatography, BDLI and BDL2 have been purified to homogeneity, and BDL3 has been partially purified to three bands on SDS/PAGE. BDLI has a molecular-mass estimate of 390 kDa by gel filtration and approx. 158 kDa by SDS/PAGE under non-reducing conditions, further reduced to subunits of 36 kDa under reducing conditions. BDL2 has a molecular mass of approx. 140 kDa and is composed of subunits of 67 kDa which can be further reduced to identical subunits of 23 kDa. Isoelectric focusing in agarose gels revealed that BDLI and BDL2 both focused as single bands at pH 6.0 and pH 5.2 respectively. The purified forms of BDLl and BDL2 were stained by the periodic acid/Schiff's reagent showing that both lectins are glycoproteins. In addition, BDL1 was deglycosylated by endo-,8-N-acetylglucosaminidase H. Immunological tests * To whom correspondence should be addressed.
Biochemical Journal, 1993
Three agglutinins (lectins), designated BDL1, BDL2 and BDL3, were identified in the haemolymph of the cockroach Blaberus discoidalis by erythrocyte cross-adsorption and sugar inhibition tests. With the use of (NH4)2SO4 fractionation, anion-exchange and affinity chromatography, BDL1 and BDL2 have been purified to homogeneity, and BDL3 has been partially purified to three bands on SDS/PAGE. BDL1 has a molecular-mass estimate of 390 kDa by gel filtration and approx. 158 kDa by SDS/PAGE under non-reducing conditions, further reduced to subunits of 36 kDa under reducing conditions. BDL2 has a molecular mass of approx. 140 kDa and is composed of subunits of 67 kDa which can be further reduced to identical subunits of 23 kDa. Isoelectric focusing in agarose gels revealed that BDL1 and BDL2 both focused as single bands at pH 6.0 and pH 5.2 respectively. The purified forms of BDL1 and BDL2 were stained by the periodic acid/Schiff's reagent showing that both lectins are glycoproteins. In ...
In vitro release of lectins by Phallusia mamillata hemocytes
Developmental & Comparative Immunology, 1991
DAbstract-o:-Lactose specific lectins are re phagocytic functions (7). A possible as leased from Phallusia mamillata hemocytes sociation between cellular lectin-like ac during short-term cultures. The molecular tivity and recognition processes in the weight of the subunits, the immunological immune response has been suggested cross-reaction and the sugar specificity suggest (7). Hemocytes of several invertebrates that the released lectins are similar to those contain (3) and also presumably secrete isolated from the sonicated hemocytes. Because lectins (8,9). lectin release appears to take place indepen Tunicates possess lectins in their dently of active protein synthesis, the possibil ity exists that lectins are pre-formed, stored in blood and the humorallectins studied so hemocytes and released when in vitro condi far are specific for sialic acid and more tions stimulate the cells. frequently, for D-galactosyl residues (10 12). D-galactosyl specific lectins can be DKeywords-Tunicates; Release; Lectins. present on the hemocyte surfaces of As cidia malaca (4) and lactose specific lec tins on Phallusia mamillata hemocytes
2020
The partial purification and characterization of haemagglutinin (lectin) were carried out from the hemolymph of the adult American cockroach, Periplaneta americana. The hemolymph was drawn from cockroach and lectin was purified by a single-step method, using ammonium sulfate (NH4)2SO4 salt fractionation and gel filtration. Gel filtration showed two peaks. The Hemagglutination Activity (HA) was observed in the 20th fraction of the second peak. The purified lectin showed a molecular weight of 26.8kDa on Sodium Dodecyl Sulfate-Poly Acrylamide Gel Electrophoresis. The purified lectin showed an increase in HA at pH 7.5 and, subsequently, a sharp decline at pH 8. This indicates that HA was specific to a certain pH level. Similarly, an increase in HA was observed until 30°C, followed by a decline at 40°C. This indicates the heat labile nature of lectin. The HA showed a higher specificity to divalent Ca2+ and showed no specificity for Ba2+. It also showed a higher inhibition for sugar D-gal...