Classes and crossreactivity of proteinases in the excretory–secretory products of Caenorhabditis elegans

Abstract

Proteinases released during the in vitro maintenance of asynchronous cultures of the free-living nematode Caenorhabditis elegans were characterized on the basis of subunit composition, fluorogenic substrate specificity, inhibitor sensitivity and pH optima. Cysteine proteinases are present in the excretory-secretory products (ESP) as indicated by the hydrolysis of cathepsin fluorogenic substrates and confirmed by immunoblotting. Serine proteinases were predominant as indicated by substrate gel analysis and inhibitor studies. The presence of metallo-proteinases was also indicated by inhibitor studies. The optimal pH value for cysteine proteinases was 5.5, while serine proteinases were optimal at pH 8.0. As a control, cultures of Escherichia coli, the diet of C. elegans, were extracted separately and gave no evidence of overlap with C. elegans ESP. Cross reactivity between the ESP of C. elegans and antibodies raised against the ESP of the equine parasite Strongylus vulgaris indicated antigenic relatedness of a proteic epitope. This is the first study to characterize the ESP of C. elegans and to display its relatedness with that of S. vulgaris.