Article

ACL-I, a lectin from the marine sponge Axinella corrugata: Isolation, characterization and chemotactic activity

Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Programa de Pós-Graduação em Botânica, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Fundação Zoobotânica do Rio Grande do Sul, Porto Alegre, RS, Brazil; Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, 90035-003, Porto Alegre, RS, Brazil; Laboratório de Farmacognosia, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology DOI:10.1016/j.cbpc.2008.03.003 pp.23-30

ABSTRACT 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 Ca2+, Mg2+ and Mn2+, 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.

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Keywords

ACL-I displays chemotactic effect
 
affinity chromatography
 
denaturant agents
 
disulphide bridges
 
dog erythrocytes
 
enzymatic proteolysis
 
lectin
 
lectin activity decreases
 
lectin agglutinates native rabbit
 
marine sponge Axinella corrugata
 
optimum pH
 
pI
 
rabbit erythrocytic stroma
 
rat neutrophil
 
SDS-PAGE
 
Superose 12 HR column
 
Ultrogel AcA 44 column