Complement factor C5 in Atlantic salmon (Salmo salar): Characterization of cDNA, protein and glycosylation

Abstract

Complement component 5 (C5) is an essential factor of the defensive complement system in all vertebrates. We report the characterization of C5 cDNA and protein from Atlantic salmon (Salmo salar), a teleost fish species of high importance in aquaculture. The C5 cDNA cloned from liver is 5079 nucleotides long, whose translation product has a molecular weight of 190 kDa, with the classical β-α orientation and motifs/sites for β-α cleavage (⁶⁷⁸RPKR⁶⁸¹) and cleavage by C5 convertases (R⁷⁵⁸). Mass spectrometric analysis show a single N-linked, biantennary, complex glycan at N¹¹²⁵. Moreover, the N-linked glycan displays an unusual modification in the form of acetylated sialic acid residues. Three anti-C5 antisera produced in mice using purified C5 worked in immunohistochemical analyses of formalin fixed liver tissue. The purification method, whereby inactive and activated (C5b) forms were isolated, opens for interesting studies on the complement function in fish, including possible connection to stress, disease and glycosylation.

Keywords: Antibodies; Atlantic salmon (Salmo salar); Complement factor C5; Glycosylation; Protein characterization; cDNA cloning.