Cathepsin B cysteine protease enzymes have been shown to be involved in a variety of different biological processes in eukaryotes. We have isolated and characterized four distinct cathepsin B-like genes from the genetically tractable nematode, Caenorhabditis elegans. This is the first reported finding of a cathepsin B-like multigene family within a nonparasitic metazoan. The four genes possess distinct genomic architectures, with variations in the position, number, and size of introns. The predicted amino acid sequences of the four genes are highly diverged. Phylogenetic analysis indicates the divergence of this multigene family within C. elegans is as great as the interspecies divergence between the vertebrates and nematode cathepsin B-like genes. In addition, each of the four genes described here shows a distinct temporal pattern of expression during C. elegans development.