Lectinas da esponja marinha Haliclona (Soestella) caerulea

Abstract

Lectins are proteins/glycoproteins that recognize carbohydrate of a specific way, but not participate in the metabolism of the same and do not belong to any of major classes of immunoglobulins. Lectins are ubiquitous proteins, present in all known organisms. In animal cells, lectins have been found in the cytoplasm, in the nucleous and as membrane-associated proteins, in diverse organelles and cells. Animal lectins can be classified into distinct families based on their physicochemical characteristics, especially in their function and identity of primary and tertiary structure. The aim of this study was to purify, characterize structural and biologically new lectins from the marine sponge Haliclona (Soestella) caerulea. H. caerulea specimens were collected in Paracuru beach, Ceará. Two lectins (H-1 and H-3) were isolated by classical techniques of protein chemistry. The primary structure of H-3 was determined by mass spectrometry and RACE. The toxic activity of lectins was evaluated against Artemia nauplii and Escherichia coli and Staphylococcus aureus strains. H-1 and H-3 showed distinct characteristics of the lectin previously isolated from H. caerulea. H-1 is a monomeric protein of 40 kDa whereas H-3 is a heterogeneus protein with chains of 9, 16 and 18 kDa. H-3 binds human erythrocytes of A and B type and was inhibited by GalNAc and PSM, H-1 binds different blood groups and could not be inhibited by any sugar tested. H-1 was toxic to Artemia nauplii (LC50 = 6.4 μg.mL-1) and H-3 was not considered toxic (LC50 = 414.2 μg.mL-1). H-3 is a blue protein that interacts with a chromophore of 597 Da of maximum absorbance at 620 nm. The primary structure of H-3 revealed a unique amino acid sequence no similar to any animal lectins known. H-3 has a hybrid glycan comprising by Hex7NAcHex7DeoxiHex2. The α-chain of H-3 undergoes complex proteolytic processing that not been fully elucidated. Moreover, H-3 was crystallized, but was not possible to obtain a diffraction pattern that permits solving the structure. In short, two new lectins were isolated and out first observed the interaction between a lectin and natural chromophore. Furthermore, for the first time given the composition glicidic out of a sponge lectin.