Echinacea purpurea (L.) Moench (EP) is a well-studied plant used for health benefits. Even though there are a lot of data on EP secondary metabolites, its active proteins are not studied well enough. The aim of our experiment was to purify lectin fraction from EP roots and evaluate its biological activity in vitro as well as its effect on kidney morphology in vivo. An EP root glycoprotein fraction was purified by affinity chromatography, identified by LC-MS/MS, and used for biological activity tests in vitro and in vivo. Identified glycoproteins were homologous with the LysM domain containing lectins from the Asteraceae plants Helianthus annuus L., Lactuca sativa L., Cynara cardunculus L. A purified fraction was tested by hemagglutination and hemagglutination inhibition (by carbohydrate reactions) in vitro. We purified the hemagglutinating active ~40 kDa size lactose, D-mannose, and D-galactose specific glycoproteins with two peptidoglycan binding LysM (lysine motif) domains. Purified LysM lectin was tested in vivo. Eight-week old Balb/C male mice (n = 15) were treated with 5 μg of the purified lectin. Injections were repeated four times per week. At the fifth experimental week, animals were sedated with carbon dioxide, then euthanized by cervical dislocation and their kidney samples were collected. Morphological changes were evaluated in hematoxylin and eosin stained kidney samples. The purified LysM lectin induced a statistically significant (p < 0.05) kidney glomerular vacuolization and kidney tubular necrosis (p < 0.001).
Fractionation and evaluation of proteins in roots ofEchinacea purpurea (L.) MoenchEchinacea purpurea (L.) Moench, a member of the Asteraceae family, is a plant rich in flavonoids, essential oils, phenolic compounds, saponins, polysaccharides and glycoproteins. The aim of the study was to evaluate the protein content in dried roots of Echinacea purpurea (L.) Moench after homo genization of roots with liquid nitrogen, extraction in 0.01 mol L -1 phosphatebuffered saline (PBS) and purification followed by fractionation of proteins using gel filtration chromatography. Total concentration of proteins was measured using the Bradford method, and evaluation of the molecular mass of proteins was accomplished by applying the SDSPAGE gel electrophoresis. The Bradford assay revealed that the highest concentration of proteins in fractions collected after gel filtration chomatography was 4.66-6.07 mg mL -1. Glycoproteins, alkamides and polysaccharides in roots of Echinacea purpurea (L.) Moench are chemical compounds that are responsible for their immunomodulatory properties. However, information about the difference of protein contents in fresh and dried roots of E. purpurea is insufficient. Keywords: Echinacea purpurea (L.) Moench, proteins, gel filtration chromatography, SDSPAGEEchinacea purpurea (L.) Moench (Asteraceae) is a herb well known for its usage in medicine. It is rich in phenolic compounds, including caffeic acid derivatives (0.6-2.1 %) -mainly cichoric, caftaric, chlorogenic and isochlorogenic acids and flavonoids (e.g., rutoside, quercetin and kaempferol). Alkamides (0.01-0.04 %) mostly isobutylamides of C11-C16 straightchain fatty acids with olefinic or acetylenic bonds are also characteristic constituents of the roots of E. purpurea. Polysaccharides (inulin, arabinogalactan), proteins and glycoproteins extracted from the roots of E. purpurea exhibit antiinflammatory activity by stimulating the immune cells. Other constituents, including the saturated pyrrolizidine type alkaloids, such as tussilagine andisotussilagine, polyacetylene derivates, and up to 0.2 % of essential oils (e.g., containing caryophyllene or humulene, limonene and camphene) have also been isolated from the roots of E. purpurea (1). Even though plant proteins
Urtica dioica L is a plant rich in flavonoids, carotenoids, caffeoylmalic acid and has an established medical value. Although content of mineral and organic substances of U. dioica L. herb is well characterized, presence of bioactive polypeptides is much less appreciated. Seeds and roots of nettle, have been established as a common source for isolation of lectins. Therefore data on the presence of lectins in herb of nettle is ambiguous. Lectin-enriched protein fractions were isolated from herb (fresh and dry) and dry extract of U. dioica L. by using homogenisation with fluid nitrogen, extraction in 0.01 M phosphate-buffer saline (PBS), concentrating, salting and precipitation. The amount of protein was measured using photometric Bradford method. A proteomic analysis using 2D gel electrophoresis was performed for lectinenriched protein fractions isolation and analysis. We estimated quantity of protein and lectins, assessed their blood cell agglutinating activity using tests employing rabbit erythrocytes. The highest concentration of protein and specific hemagglutination activity was observed for protein fractions isolated from fresh herb. The highest lectins content was presented in protein fractions isolated from the dry extract.
Proteins were extracted and fractionated from Echinacea purpurea L. (Moench) (purple coneflower) roots, to evaluate their hemagglutinating activity. Our data suggests that the ~ 25 kDa glycosylated protein is responsible for the hemagglutinating activity and might be worth for its investigation for other biological activities.
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