title: Molecular cloning and primary sequence of a cysteine protease expressed by Haemonchus contortus adult worms.
abstract: We have cloned cDNAs encoding a 35-kilodalton cysteine protease that is a major component of protective extracts isolated from blood-feeding Haemonchus contortus adult worms. Near full-length cDNAs for the protease were isolated by immunoscreening an adult worm cDNA expression library with a rabbit antiserum prepared against the protein eluted from preparative SDS gels and by rescreening the library with oligonucleotide probes. The protein predicted from the nucleotide sequence of the cDNAs and of a genomic DNA clone comprises 342 amino acids and contains an N-terminal signal sequence, 16 cysteine residues and four potential N-linked glycosylation sites. The enzyme appears to be glycosylated in vivo. The H. contortus protease, called AC-1, displays an overall 42% sequence identity with the human lysosomal thiol protease cathepsin B. The similarities between cathepsin B and AC-1 are localized primarily to regions of cathepsin B that comprise the mature, active form of the enzyme. A stretch of six amino acids that includes the active site cysteine of cathepsin B is conserved, and is present in the same relative location in AC-1, suggesting that this region comprises the active site of the H. contortus enzyme.
authors.affiliation: Synergen, Inc., Boulder, CO 80301.
authors.forename: G N
authors.initials: GN
authors.lastname: Cox
authors.forename: D
authors.initials: D
authors.lastname: Pratt
authors.forename: R
authors.initials: R
authors.lastname: Hageman
authors.forename: R J
authors.initials: RJ
authors.lastname: Boisvenue
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics*; D004247:DNA / Q000235:genetics; D006031:Glycosylation; D006189:Haemonchus / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014251:Trichostrongyloidea / Q000201:enzymology*
chemical_list: D004247:DNA; D003546:Cysteine Endopeptidases