S-type lectins occur also in invertebrates: High conservation of the carbohydrate recognition domain in the lectin genes from the marine sponge Geodia cydonium

Abstract

The marine sponge Geodia cydonium contains several lectins. The main component, called lectin-1, is composed of three to four identical subunits. The subunits of the lectins were cloned from a cDNA library; two clones were obtained. From the deduced aa sequence of one clone, LECT-1, a mol. wt of 15,313 Da is calculated; this value is in good agreement with mass spectrometric analysis of 15,453 +/- 25 Da. The sequence of another clone, LECT-2, was analysed and the aa sequence was deduced (15,433 Da). The two subunits have a framework sequence of 38 conserved aa which are characteristic for the carbohydrate-binding site of vertebrate S-type lectins. Clustering of lectin sequences of various species following their pairwise comparison establishes a dendrogram, which reveals that the sponge lectin could be considered as the ancestor for vertebrate S-type lectins.

Expression of the potential biomarker heat shock protein 70 and its regulator, the metazoan DnaJ homolog, by temperature stress in the sponge Geodia cydonium

“…'treated samples '. Polymerase chain reaction-cloning of GCDNAJ from Geodia cydonium. The N-terminus of the cDNA encoding sponge DnaJ-like protein was cloned by poly-merase chain reaction (PCR) from a G. cydonium cDNA library (Pfeifer et al 1993) in lambda ZAPII, using the degenerate reverse primer 5'-RTCIGGRT-GRTAHAIICTIGC-3' (where I = inosine; H = C/T and R = AIG) in conjunction with the ZAPII 5'-end vectorspecific primer T3. The PCR reaction mixtures of 50 p1 included 10 pm01 of the degenerate primer and 5 pm01 of the primer T3 (Stratagene), 200 FM of each nucleotide, 1 p1 of the cDNA library [approximately 10" plaque-forming units (pfu)], buffer and 2.5 units of Taq DNA polymerase (Boehringer Mannheim).…”

Sponge Bcl-2 homologous protein (BHP2-GC) confers distinct stress resistance to human HEK-293 cells

“…This cDNA was used for expression in Escherichia coli as described. 53,69 The cDNA was inserted into the bacterial oligohistidine expression vector pQE-32 (Quiagen). E. coli (XL1-blue) were transformed with this plasmid and expression of fusion protein was induced with isopropyl 1-thio-b-Dgalactopyranoside (IPTG).…”

Molecular and functional characterization of a tandem-repeat galectin from the freshwater snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni

“…These are most highly represented in such organisms as nematodes (Hirabayshi et al, 1992;Greenhalgh et al 1999;Newlands et al, 1999), arthropods (Pace et al, 2002;Baum, 2004, Barat-Houari et al, 2006;Huang et al, 2007;Kamhawi et al 2004), tunicates (Parrinello et al, 2007) and sponges (Pfeifer et al, 1993;Stalz et al, 2006). In the Phylum Mollusca, the presence of galactose-binding lectins also has been demonstrated (e.g., Suzuki and Mori, 1989;Mitra and Sakar, 1998;Wilson et al, 1992;Ozeki, 1998), and in some species, the molecular mass of isolated lectins were consistent with galectins possessing single (Mitra and Sakar, 1998) or dual (Ozeki, 1998) CRDs.…”