ACL-I, a lectin from the marine sponge Axinella corrugata: Isolation, characterization and chemotactic activity Roger R. Dresch a , Gilberto D. Zanetti b , Cléa B. Lerner c , Beatriz Mothes c , Vera M.T. Trindade d , Amélia T. Henriques e , Magdolna M. Vozári-Hampe d, ⁎ a Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil b Programa de Pós-Graduação em Botânica, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil c Fundação Zoobotânica do Rio Grande do Sul, Porto Alegre, RS, Brazil d Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, 90035-003, Porto Alegre, RS, Brazil e Laboratório de Farmacognosia, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil ABSTRACT ARTICLE INFO Article history: Received 7 February 2008 Received in revised form 5 March 2008 Accepted 5 March 2008 Available online 12 March 2008 Keywords: Axinella corrugata Chemotaxis Lectin Marine sponge The lectin from the marine sponge Axinella corrugata (ACL-I) was purified by affinity chromatography on rabbit erythrocytic stroma incorporated into a polyacrylamide gel followed by gel filtration on Ultrogel AcA 44 column. Purified ACL-I is a hexameric glycoprotein with a Mr of 82.3 kDa estimated by SDS-PAGE and 78.5 kDa by FPLC on Superose 12 HR column. The pI of lectin is 6.3 and ACL-I is constituted of 13.9 kDa similar subunits some of them linked by disulphide bridges. This lectin agglutinates native rabbit, goat and dog erythrocytes and in less extent human erythrocytes. The hemagglutinating activity is independent of Ca 2+ , Mg 2+ and Mn 2+ , but it is strongly inhibited by carbohydrates containing N-acetyl groups. ACL-I is stable up to 70 °C for 30 min, with optimum pH between 7 and 8, and it is also resistent to enzymatic proteolysis in vitro. In the presence of reducing or denaturant agents, the lectin activity decreases. ACL-I displays chemotactic effect on rat neutrophil in vitro which is inhibited by N-acetyl-D-glucosamine. © 2008 Elsevier Inc. All rights reserved. 1. Introduction Sponges (Phylum Porifera) are among the oldest metazoan animals and are the simplest of the multicellular animals. Because of their prevalence, distribution and ability to biosynthesize a variety of classes of natural compounds, the sponges have become one of the dominant sources of biologically active marine natural products. Among these compounds are the lectins (Carté, 1996; Hentschel et al., 2006). Lectins are (glyco)proteins that possess at least one noncatalytic domain able to recognize and to bind, in a reversible manner and with certain specificity, carbohydrates free or complexed, without modify- ing them (Peumans and van Damme, 1995). Lectins show considerable differences in their proteic structures, characteristics and conse- quently in their biological properties. Because of their capacity to detect subtle differences among complex carbohydrate structures binding to them and their high stability (relatively good resistance to acidic pH and degradation by gastrointestinal proteolytic enzymes), lectins are valuable tools for detection, isolation and characterization of glycoconjugates, for cyto and histochemistry, for the examination of changes that occur on cell surfaces during physiological and pathological processes, from cell differentiation to cancer and for the study of immunological and inflammatory processes (Zatta and Cummings, 1992; Ponchel and Irache,1998; Ezpeleta et al.,1999; Dias-Baruffi et al., 2000; Gabius et al., 2002; Sharon and Lis, 2004; Sharon, 2007). Although lectins are largely found in nature and have countless biological and medical applications, their physiological roles in marine invertebrates and especially in marine sponges are not well known. The lectin from the marine sponge Cliona varians was described and displays cytotoxic effect on Gram positive bacteria and agglutinates Leishmania promastigotes, indicating physiological defense roles of the protein (Moura et al., 2006). The lectin of the sponge Haliclona cratera also showed cytotoxicity towards HeLa and FemX cells (Pajic et al., 2002). Furthermore, this lectin displays mitogenic effect in vitro on human T lymphocytes, as well as Axinella polypoides lectin I (Phillips et al., 1976). Mitogenic activity was also a characteristic of the lectin isolated from the sponge Craniella australiensis (Xiong et al., 2006). On the other hand, one feature of Geodia cydonium lectin I is to link the aggregation factor to cells in the adhesion systems of the sponge (Wagner-Hülsmann et al., 1996). During the screening for lectin activity in aquous extracts from marine sponges of Brazilian South Atlantic Coast with the aim to search for new lectins with distinct structures and properties (Dresch et al., 2005), we found that the extracts from Axinella corrugata strongly agglutinated rabbit erythrocytes. In the present work we Comparative Biochemistry and Physiology, Part C 148 (2008) 23–30 ⁎ Corresponding author. Tel.: +55 51 33085561; fax: +55 51 33085534. E-mail address: rogdresch@gmail.com (M.M. Vozári-Hampe). 1532-0456/$ – see front matter © 2008 Elsevier Inc. All rights reserved. doi:10.1016/j.cbpc.2008.03.003 Contents lists available at ScienceDirect Comparative Biochemistry and Physiology, Part C journal homepage: www.elsevier.com/locate/cbpc