: Lectins are class of proteins characterized by their ability to selectively bind carbohydrate moieties of glycoproteins. Many invertebrate lectins, especially derived from hemolymph are being purified and yet their functions and medical applications are subjects of major interest. Hemolymph lectins in invertebrates play a major role in protecting against many pathogens and microbes. Further, many hemolymph lectins show anticancer properties towards various cancer cell lines which expresses globotriaosyl ceramides on their cell surface. These vast repertoires of hemolymph lectins in recognizing and inhibiting the growth of various harmful microbes and cancerous cells have spurred the biochemist to use them in histochemical and cytochemical studies. The present review will address the biological roles and biomedical applications of hemolymph lectin.
A new thermostable caseinolytic serine protease was purified from the latex of Euphorbia heterophylla L. to electrophoretic homogeneity by a procedure involving successive steps of pretreatment of the latex, PEG fractionation, CM-cellulose chromatography and DEAE-cellulose chromatography. The purified protease was found to be a monomeric protein of molecular weight 77.2 kDa. It exhibited caseinolytic activity with hyperbolic azocasein saturation with Vmax and Km values of 0.11 units.mL(-1) and 0.55 mg.mL(-1) respectively. Specific inhibitory studies revealed the enzyme to be a serine protease. The protease was characterized by pH optimum of 8.0 and high thermostability with T1/2 of 75°C. Based on the results of peptide mass fingerprinting analysis, the protease was shown to be a new protein not characterized earlier.
Background: Lectins are proteins or glycoproteins of non-immune origin which bind specifically but reversibly to carbohydrates or glycoconjugates. They play a crucial role in various biological processes including host defense mechanism, inflammation and metastasis. Therefore, there is an expanding scientific emphasis on purification and characterization of novel lectins possessing different useful biological properties. Objective: The present investigation is concerned with purification and characterization of a novel lectin from the hemolymph of oak tasar (Antheraea proylei J.) silkworm. Methods: The lectin was purified from the hemolymph by a procedure involving successive steps of hemocyte-free hemolymph preparation, ammonium sulfate (0-40%) fractionation and affinity chromatography on a column of Sephadex G-50 covalently coupled with L-rhamnose. It was then characterized by various physic-chemical methods including SDS-PAGE, gel filtration, hemagglutination assay, hemagglutination inhibition assay and tandem mass spectrometry (LC-MS/MS) coupled with Mascot sequence matching software (Matrix Science). Results: The lectin was purified to electrophoretic homogeneity from the silkworm hemolymph and was found to be a monomeric protein with a native molecular weight of 39.5 kDa. It was specifically inhibited by L-rhamnose and D-fucose, the former being sixteen times more inhibitory than the latter. The hemagglutinating activity further was characterized by independency of metal ion, optimum at pH 7-7.5 and thermal stability with t1/2 of 60°C. Analysis with tandem mass spectrometry coupled with Mascot sequence matching software confirmed the purified lectin to be a protein not purified and characterized earlier. Conclusion: A novel rhamnose/fucose-specific lectin was purified to electrophoretic homogeneity from hemolymph of oak tasar (Antheraea proylei J.) silkworm. The lectin was found to be a monomeric protein with a native molecular weight of 39.5 kDa. Its activity was found to be independent of metal ion, optimum at pH 7-7.5 and characterized by thermal stability with t1/2 of 60°C. Analysis with tandem mass spectrometry coupled with Mascot sequence matching software confirmed the purified lectin to be a protein not characterized earlier.
Plant lectins are carbohydrate binding proteins that are ubiquitously found in almost all plant species and have different structures and functions depending on the sources. Purification of lectins from their plant sources and determination of their sugar specificity become an important goal for the evaluation of their potential biomedical applications. Here, we report the affinity purification of a D-galactose specific lectin from the seeds of Meizotropis buteiformis Voight., and its physicochemical parameters, and LC-MS/MS (tandem mass spectrometry) analysis. Isolation and purification of this lectin were performed by simple successive steps of lectin extraction, ammonium sulphate fractionation, and affinity chromatography using lactose-linked Sepharose-4B chromatography column. The affinity-purified lectin has a native molecular weight of 75 kDa and is found to be a heterodimer (molecular weight of 36 and 38 kDa). The LC-MS/MS results suggested that the purified lectin has not been reported earlier. Aim: The main aim of the present study is to find out the novelty and characteristics of a lectin purified from the plant Meizotropis buteiformis. Background: Lectins are proteins ¬¬that possess the ability to specifically bind glycans of glycoconjugates. Plants are considered rich sources of lectins and determination of sugar specificity of a purified plant lectin is an important aspect in order to evaluate its potential area of application. In the present study, a novel D-Galactose specific lectin is purified from Meizotropis buteiformis through affinity chromatography and examined for its various physical and biochemical characteristics. Objective: Objective of the present study is to purify a novel lectin upto its homogeneity from the seeds of Meizotropis buteiformis and characterization of its various physical and biochemical properties. Methods: The lectin was purified by simple successive steps of lectin extraction, ammonium sulphate fractionation, and affinity chromatography. Activity of the purified lectin was determined by hemagglutination assay. Some physicochemical parameters of the purified protein were also determined along with identification of protein by LC-MS/MS and the spectra analysis using Mascot sequence matching software (Matrix Science) with NCBI database. Results: From the current investigation, it was found that the purified lectin has a native molecular weight of 75 kDa. Among the various sugars and sugar derivatives tested, lactose and D-galactose were found to be potent inhibitors of its activity. Its optimum pH range was found to be from 6.5 to 7.5 and also it exhibited full activity at temperature from 00C to 500¬C. The purified lectin does not show any effects on its activities for metal ions tested and protein view report of the LC-MS/MS result analysis showed a 50% sequence similarity with that of lectin beta-chain of the Butea monosperma. Conclusion: In the present study, a novel D-Galactose specific lectin is purified from Meizotropis buteiformis by affinity chromatography using Sepharose 4B. The purified lectin is found to be a heterodimeric and metal ion independent. The LC-MS/MS results suggested that the purified lectin has not been reported earlier.
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