Lectins In Extracts Research Articles

LECTINS ARE INVOLVED IN THE RECOGNITION BETWEEN ANABAENA AND AZOLLA

ABSTRACT The presence of a lectin in extracts of the endosymbiotic Anabaena azollae was demonstrated using the haemagglutination assay. This activity was sensitive to D(—) fucose and L(—) rhamnose but was not affected by other saccharides tested. Furthermore, an endogenous inhibitor was extracted from isolated leaf cavity envelopes of Azolla, suggesting the possibility of a recognition site for the lectin in the host.

A new assay for functional lectins: the brush border lectin agglutination assay (BBLAA)

A new method, the brush border lectin agglutination assay (BBLAA), has been developed to measure the capacity of functional lectins to agglutinate the intestinal brush border membrane. Lectins were incubated with purified chick small intestinal brush border vesicles, agglutinated membrane was isolated by low speed centrifugation and then assayed using alkaline phosphatase as a marker. The brush border agglutination activity of purified plant lectins with various carbohydrate binding specificity was compared. Tomato ( Lycopersicon esculentum) lectin, and wheat germ ( Triticum vulgaris) agglutinin bind N-acetyl- d-glucosamine and were found to have the highest activity. An exponential sigmoid equation fit the data obtained with wheat germ agglutinin, which is consistent with a cooperative mechanism of brush border agglutination. The N-acetyl- d-galactosamine specific lectin from soybean ( Glycine max), and the l-fucose specific lectin from asparagus pea ( Lotus tetragonolobus) had intermediate activity. Of the mixed carbohydrate binding isolectins from red kidney bean ( Phaseolus vulgaris), Pha-E had intermediate activity, and Pha-L no detectable activity. Jack bean ( Canavalia ensiformus) lectin, concanavalin A, (Con A) binds glucose/mannose residues and had marginal activity. Succinylation of con A eliminated brush border agglutination activity. The BBLAA has distinct advantages when compared to other lectin assay systems in that it measures functional agglutination activity directed toward the target membrane on the intestinal epithelium, does not require antibody or enzyme conjugation, and can be used to measure total functional lectins in extracts from diets or meals.

Inhibitory action of spray dried blood plasma and whole egg powder on lectins in extracts of several legume seeds: a qualitative approach

Samples of seeds from eight legume species and Triticum vulgaris grains were extracted with buffer and lectin activity in the extracts was determined in haemagglutination experiments using normal or Pronase-treated rabbit erythrocytes. The effect of the addition of spray dried porcine and bovine plasma powder, whole egg powder, galactosides, whey powder and specific inhibitors (eg mannose, galactose, N-acetyl galactosamine, fetuin) on haemagglutination activity (HA) was determined. Plasma powders were potent inhibitors of HA in extracts of Pisum sativum, Vicia faba, Vicia sativa, Lens culinaris and Phaseolus vulgaris. HA in extracts of Lupinus sp and Phaseolus vulgaris was efficiently decreased by whole egg powder, while the lectin of Glycine max could only be inhibited by addition of galactosides, whole and defatted milk powder and whey powder. Inhibitors (plasma and whole egg powder and fetuin) were subjected to SDS-PAGE and Western blotting and blots were incubated with biotinylated lectins, except for Lupinus lectin. Results of the HA experiments were confirmed: lectins which were not influenced by inhibitory compounds in HA experiments also showed no binding with proteins of the blotted inhibitor. There were strong indications that lectins were not bound to the albumin fraction of the plasma powders. Results are discussed in view of future in vivo experiments. © 1998 SCI.

Inhibitory action of spray dried blood plasma and whole egg powder on lectins in extracts of several legume seeds: a qualitative approach

Inhibitory action of spray dried blood plasma and whole egg powder on lectins in extracts of several legume seeds: a qualitative approach

Expression of a 31‐kDa lactoside‐binding lectin in normal human gastric Mucosa and in primary and metastatic gastric carcinomas

The expression of lactoside-binding lectin L-31 was analyzed in normal mucosa and in primary and metastatic gastric carcinomas. Immunoblotting revealed L-31 lectin in extracts of normal and malignant gastric tissues from 26 patients. The L-31 level was higher in tumor than in normal tissue in 9/26 cases, similar in 14/26 cases, and lower in 3/26 cases. Anti-L-31 monoclonal antibodies (MAbs) were used in immunohistochemical analyses to compare lectin expression in specimens of primary gastric carcinomas and adjacent normal mucosa from 39 patients and in specimens of metastases and the corresponding primary gastric carcinomas from 74 patients. The lectin was detected in normal gastric epithelial cells and in all gastric carcinoma specimens, albeit in varying amounts. The L-31 level was significantly higher in the primary tumor than in adjacent normal tissue in 55% of the well-differentiated tubular carcinoma cases and in 50% of stage-III and -IV tumors. L-31 expression in liver metastases from well-differentiated tubular primary gastric carcinomas was higher in 31% of the cases relative to the corresponding primary cancers. Likewise, L-31 expression in metastases from poorly differentiated gastric carcinomas in lymph nodes was higher in 38% of the cases compared to the primary cancers. The higher expression of L-31 in primary cancers and metastases of certain types implicates this lectin in the metastatic phenotype, but the presence of L-31 in a primary cancer is not sufficient to allow the metastatic propensity of the tumor to be predicted.

Lectins in Castor Bean Seedlings

The amounts of the two lectins (ricin and Ricinus communis agglutinin) in tissues of castor bean seedlings were followed during germination and early growth. For measurement, lectins in extracts were separately eluted from Sepharose columns; an antibody to the agglutinin was also used to detect the lectins by immunodiffusion. The endosperm of the dry seed contains 3.5 mg total lectin (5.6% of the total seed protein), which declines by 50% by day 4 and more rapidly thereafter as the tissue is completely consumed. The cotyledons of the dry seed also contain lectins but the amounts are less than 1% of those in the endosperm, and, as in the endosperm, they are constituents of the albumin fraction of the isolated protein bodies. No lectins were detected in the green cotyledons of 10-day seedlings that had been exposed to light from day 5. The embryonic axes of 2-day seedlings contained very small amounts of lectins but they were not detectable in the aerial parts of seedlings grown for 3 weeks or in cells from endosperm grown in tissue culture.The ability of proteinases and glycosidases (isolated from endosperm of 4-day seedlings) to hydrolyze the lectins was examined. No hydrolysis of the two lectins was observed, but the subunits, separated by reduction with 2-mercaptoethanol, were hydrolyzed slowly by a proteinase and some release of mannose was observed in the presence of the glycosidases. Ricin was converted to its subunits by cysteine and an enzyme in an endosperm extract accelerated chain separation by glutathione.

Distribution of Lectins in Tissues, Derived Callus, and Roots of Psophocarpus tetragonolobus (Winged Bean)

The distribution of lectin in parental tissues, roots formed de novo from parental stem tissue, and derived callus cells of Psophocarpus tetragonolobus has been measured by hemagglutinating activity and radioimmunoassay. The antisera used for the radioimmunoassay was raised in rabbits to lectin isolated from seeds by affinity chromatography using insolubilized hog gastric mucin. The distribution of lectin in buffer extracts of the tissues (or cells) and the extracellular medium favors the tissues for in vitro grown roots, regardless of the culture conditions used. The lectin content of the extracellular medium is more significant for callus, regardless of its conditions of culture. The lectin activity of extracts of in vitro grown roots was higher than that of mature roots from whole plants. Differences in relative levels of lectin activity measured by hemagglutination and by radioimmunoassay, and differences in saccharide inhibition of hemagglutination, suggest the presence of multiple lectins in extracts of different tissues.

Developmentally regulated lectins from chick muscle, brain, and liver have similar chemical and immunological properties

Developmentally regulated lectins in extracts from brain, liver and muscle of 16-day-old chick embryos and liver of 7-day-old chicks have been purified by affinity chromatography. The purified preparations from the different tissues were indistinguishable in molecular weight and isoelectric point. The lectins could also not be distinguished when tested as antigens with antiserum raised against highly purified muscle lectin. This apparent identity was indicated both in double gel diffusion tests and by determination of the antibody-mediated inhibition of hemagglutination activity of the various lectins. Thiodigalactoside and lactose were potent inhibitors of the lectins from all sources. Galactose was a less potent inhibitor, especially with preparations from embryonic liver. After isoelectric focusing of these purified preparations, they all showed reduced and equivalent galactose sensitivity. Since the lectins from the different tissues appear identical, there is presently no basis to infer that they impart qualitative uniqueness to these tissues during differentiation.

Lectins in extracts of certain Polygonaceae seed precipitate animal and human serums.

Seeds of four species of Polygonaceae were tested for lectins that precipitate human and animal serums. Rumex crispus, Polygonum convolvulus, and Polygonum pennsylvanicum developed specific precipitate bands on double diffusion on agar gel plates. These bands were enhanced and increased in number when extracts were tested against serums from patients with certain diseases. When tested against lyophilized serum, no precipitate bands developed. The active substance cannot be dialyzed through cellulose membrane against running tap water for 16 hours, and it is heat stable. Extracts from Fagopyrum esculentum developed no precipitate bands.