Recombinant production and characterization of the carbohydrate recognition domain from Atlantic salmon C-type lectin receptor C (SCLRC).
The Atlantic salmon C-type lectin receptor C (SCLRC) locus encodes a potential oligomeric type II receptor. C-type lectins recognize carbohydrates in a Ca(2+)-dependent manner through structurally conserved, yet functionally diverse, C-type lectin-like domains (CTLDs). Many conserved amino acids in animal CTLDs are present in SCLRC, with the notable exception of an asparagine crucially involved in Ca(2+)- and carbohydrate-binding, which is tyrosine in SCLRC. SCLRC also contains six cysteines that form three disulfide bonds. Although SCLRC was originally identified as an up-regulated transcript responding to Aeromonas salmonicida infection, the biological role of this protein is still unknown. To study the structure and ligand binding properties of SCLRC, we created a homology model of the 17kDa CTLD and produced it as an affinity-tagged protein in the periplasm of Escherichia coli by co-expression of proteins that facilitate disulfide bond formation. The recombinant form of SCLRC was characterized by a protease protection assay, a solid-phase carbohydrate-binding assay, and frontal affinity chromatography. On the basis of this characterization, we classify SCLRC as a C-type lectin that binds to mannose and its derivatives.
Soanes
Kelly H
KH
NRC Institute for Marine Biosciences, National Research Council Canada, 1411 Oxford Street, Halifax, NS, Canada B3H 3Z1.
Ewart
K Vanya
KV
Mattatall
Neil R
NR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000494:pharmacology; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D037181:Lectins, C-Type / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D020317:Salmo salar; D016415:Sequence Alignment; D014357:Trypsin / Q000378:metabolism
D037181:Lectins, C-Type; D011994:Recombinant Proteins; D014357:Trypsin; D008358:Mannose; D002118:Calcium; D005690:Galactose