Identification of <i>Echinacea Purpurea</i> (L.) Moench Root LysM Lectin with Nephrotoxic Properties

• Main Authors: Gabriele Balciunaite, Perttu-Juhani Haimi, Zoja Mikniene, Girius Savickas, Ona Ragazinskiene, Nomeda Juodziukyniene, Danas Baniulis, Dalia Pangonyte
• Format: Article
• Language: English
• Published: MDPI AG 2020-01-01
• Series: Toxins
• Subjects: <i>echinacea purpurea</i> l. moench; lectin; hemagglutinin; lysm; lectin purification; affinity chromatography; hemagglutinating activity; nephrotoxicity
• Online Access: https://www.mdpi.com/2072-6651/12/2/88

<i>Echinacea purpurea</i> (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 <i>Asteraceae</i> plants <i>Helianthus annuus</i> L., <i>Lactuca sativa</i> L., <i>Cynara cardunculus</i> 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 &#956;g 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 (<i>p</i> &lt; 0.05) kidney glomerular vacuolization and kidney tubular necrosis (<i>p</i> &lt; 0.001).