Repetitive peptide motifs in the cuticlin of Ascaris suum

The cuticle of parasitic nematodes is composed of extracellular structural proteins. Over 90% of these proteins are collagenous molecules in the basal and median layers of the cuticle. The outermost layers of the cuticle, the epicuticle, is composed of non-collagenous proteins, that represent the structural surface of nematodes. In Ascaris these proteins have been termed 'cuticlins'. While cuticular collagens have been well studied by both biochemical and genetic means, knowledge of the molecular structure of cuticlin components of parasitic nematodes is scarce. In the present paper we report on the production of monoclonal antibody 8.1, which is specific for cuticlin, but does not recognize collagen epitopes. We have screened a cDNA library derived from adult Ascaris suum mRNA of the hypodermal tissue underlying and synthesizing the cuticle. One positive cDNA clone encodes alanine-rich repetitive motifs, which are part of the insoluble cuticlin of the outermost layers of the epicuticle of Ascaris suum. This was shown in immunocytochemical experiments using specific polyclonal antisera raised against these motifs, expressed as fusion protein with glutathione S-transferase of the helminth Schistosoma japonicum. Comparison of the repetitive amino acid sequence with structural proteins of the nematode Caenorhabditis elegans and the insects Locusta migratoria and Ceratitis capitata revealed a minimal consensus motif.