title: Identification of an asparagine amidohydrolase from the filarial parasite Dirofilaria immitis.
abstract: The nematode cuticle is a complex extracellular structure which is secreted by an underlying syncytium of hypodermal cells. Recent studies have demonstrated that the cuticle of parasitic nematodes is a dynamic structure with important absorptive, secretory, and enzymatic activities. In addition, the cuticle serves as a protective barrier against the host. A 48-h third stage larval Dirofilaria immitis cDNA library was immunoscreened with sera raised against larval cuticles. One clone, L3MC4 that reacted strongly with the anti-cuticle antisera was sequenced. The composite cDNA sequence comprises 2073 bp coding for a full-length protein of 590 amino acids. GenBank analysis showed that DiAsp had significant similarity to a Caenorhabditis elegans gene-product (54% identity) and to other asparaginases at the amino acid level. Escherichia coli-expressed recombinant DiAsp (rDiAsp) catalysed the hydrolysis of asparagine to aspartate and ammonia. Antibodies raised against D. immitis larval cuticles reacted with rDiAsp in immunoblots. This is the first report of identification of a cDNA clone encoding an asparaginase enzyme from a parasitic nematode.
authors.affiliation: Heska Corporation, Fort Collins, Colorado 80525, USA.
authors.forename: N
authors.initials: N
authors.lastname: Tsuji
authors.forename: T H
authors.initials: TH
authors.lastname: Morales
authors.forename: V V
authors.initials: VV
authors.lastname: Ozols
authors.forename: A B
authors.initials: AB
authors.lastname: Carmody
authors.forename: R
authors.initials: R
authors.lastname: Chandrashekar
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001215:Asparaginase / Q000235:genetics* / Q000378:metabolism; D001216:Asparagine / Q000378:metabolism; D004183:Dirofilaria immitis / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015801:Helminth Proteins / Q000235:genetics; D007814:Larva / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D015801:Helminth Proteins; D011994:Recombinant Proteins; D001216:Asparagine; D001215:Asparaginase