A lactose specific lectin from the sponge Cinachyrella apion: Purification, characterization, N-terminal sequences alignment and agglutinating activity on Leishmania promastigotes

Abstract

Crude extract from the sponge Cinachyrella apion showed cross-reactivity with the polyclonal antibody IgG anti-CvL (Cliona varians lectin) and also a strong haemagglutinating activity towards human erythrocytes of all ABO groups. Thus, it was submitted to acetone fractionation, IgG anti-deglycosylated CvL Sepharose affinity chromatography, and Fast Protein Liquid Chromatography (FPLC-AKTA Purifier) gel filtration on a Superose 6 10/300 column to purify a novel lectin. C. apion lectin (CaL) agglutinated all types of human erythrocytes with preference for papainized type A erythrocytes. The haemagglutinating activity is independent of Ca2+, Mg2+ and Mn2+ ions, and it was strongly inhibited by the disaccharide lactose, up to a minimum concentration of 6.25 mM. CaL molecular mass, determined by FPLC-gel filtration on a Superose 12 10/300 column and SDS gel electrophoresis, was approximately 124 kDa, consisting of eight subunits of 15.5 kDa, assembled by hydrophobic interactions. The lectin was heat-stable between 0 and 60 degrees C and pH-stable. The N-terminal amino acid sequence of CaL was also determined and a blast search on amino acid sequences revealed that the protein showed similarity only with a silicatein. Leishmania chagasi promastigotes were agglutinated by CaL and this activity was abolished by lactose, indicating that lactose receptors could be presented in this parasite stage. These findings are indicative of the potential biotechnological application of CaL as diagnostic of pathogenic protozoa.

Hemagglutinating/Hemolytic activities in extracts of marine invertebrates from the Brazilian coast and isolation of two lectins from the marine sponge Cliona varians and the sea cucumber Holothuria grisea

“…Several quaternary arrangements have been reported in sponge lectins: dimeric (Pajic et al 2002), trimeric (Xiong et al 2006), tetrameric (Miarrons and Fresno 2000) and multimeric (Medeiros et al 2010) proteins linked by weak interactions or disulfide bonds were described.…”

Lectins: production and practical applications

“…Acids (e.g., acetic acid used by Naeem et al 2007), organic solvent (e.g., acetone used by Medeiros et al 2010) or salt (e.g., ammonium sulfate) can be used to precipitate lectins. Various chromatographic methods, including affinity chromatography, ionic exchange chromatography, hydrophobic interaction chromatography, and gel filtration can be utilized.…”

Isolation, Structural Elucidation, and Absolute Stereochemistry of Enigmazole A, a Cytotoxic Phosphomacrolide from the Papua New Guinea Marine Sponge Cinachyrella enigmatica

“…Cinachyrella is a genus in the Desmosponge family Tetillidae that has not been the focus of extensive chemical study in the past. Other than a few fatty acid derivatives 8 or steroids 9 that were isolated from various Cinachyrella species, only a lactose-specific lectin from Cinachyrella apion 10 and a tricyclic alkaloid from Cinachyrella sp. 11 have been described.…”