Cloning and sequence comparisons of four distinct cysteine proteases expressed by Haemonchus contortus adult worms.
Three new members of a developmentally regulated cysteine protease gene family of the parasitic nematode Haemonchus contortus have been isolated and characterized. One of the new genes, AC-3, was found to be linked in tandem to the previously characterized AC-2 gene. Nucleotide sequence analyses revealed that the first 90 amino acids of AC-3 are organized into four exons, similar to the situation in AC-2. A cDNA that appears to be a near full-length copy of the AC-3 gene was isolated using the polymerase chain reaction (PCR) technique to amplify cDNAs from adult worm poly(A)+ mRNAs. In addition to AC-3, a distinct cysteine protease cDNA, AC-4, was amplified by the same oligonucleotide primers. cDNAs encoding a fifth cysteine protease, AC-5, were isolated from an adult worm cDNA expression library using specific rabbit antisera and by PCR. Comparison of the predicted amino acid sequences of AC-3, AC-4 and AC-5 reveal that they share 64-77% identity with one another and with the previously reported AC-1 and AC-2 sequences. The amino acids surrounding the active site cysteine are highly conserved, as are the positions of other cysteine residues in the mature protein sequences. The H. contortus proteases are more similar to one another than they are to human cathepsin B (38-44% amino acid identity) or to the Sm31 cysteine protease of Schistosoma mansoni (36-40% identity). Our studies indicate that H. contortus adult worms express mRNAs for several distinct cysteine proteases. The significant primary sequence differences between the proteases suggest that they differ in their substrate specificities and precise physiological functions.
Pratt
D
D
Synergen, Inc., Boulder, CO 80301.
Armes
L G
LG
Hageman
R
R
Reynolds
V
V
Boisvenue
R J
RJ
Cox
G N
GN
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D004247:DNA; D005340:Fibrinogen / Q000378:metabolism; D015870:Gene Expression; D008040:Genetic Linkage; D006189:Haemonchus / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D015183:Restriction Mapping; D016415:Sequence Alignment
D005340:Fibrinogen; D004247:DNA; D003546:Cysteine Endopeptidases