Cloning and expression of a cDNA encoding a nonintegrin laminin-binding protein from Echinococcus granulosus with localization of the laminin-binding domain.
We have isolated a cDNA from the hydatid tapeworm, Echinococcus granulosus, encoding a protein that binds laminin. This is the first report of a helminth parasite laminin-binding protein and the first description of a cDNA encoding a laminin-binding protein from a parasite. The cDNA clone (egmo3) was isolated from an E. granulosus protoscolex cDNA expression library, and identified on the basis of sequence homology to the nonintegrin mammalian metastasis-associated 67-kDa laminin receptor (67-LR). The amino acid sequence predicted from the cDNA sequence is 268 residues long with a calculated molecular mass of 29.9 kDa. Southern blot analysis suggested that many copies of the gene may occur in the E. granulosus genome. A Northern blot revealed that the gene is expressed as a single transcript of approximately 1 kb consistent with the size of the cDNA insert. Antibodies raised to the purified protein interacted with a 30 kDa protein in whole E. granulosus protoscoleces. A Western blot of the purified and refolded recombinant protein specifically bound 125I-labelled laminin, as did a synthetic peptide derived from the inferred amino acid sequence of egmo3 which is similar in homology to peptide G, the active ligand-binding site of 67-LR. We also isolated the 3' end of the cDNA encoding the homologous protein from the closely related species, E. multilocularis. The polypeptide encoded by egmo3 also shares substantial identity with the acidic class of ribosomal proteins which are involved in protein synthesis. As such, the egmo3 protein may be multifunctional in E. granulosus, acting as a laminin-binding molecule but also playing a role in cell division and growth.
Zhang
L
L
Molecular Parasitology Unit, Australian Centre for International and Tropical Health and Nutrition, Bancroft Centre, Brisbane QLD, Australia.
Leggatt
G R
GR
Kalinna
B H
BH
Piva
T J
TJ
McManus
D P
DP
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics*; D004446:Echinococcus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011498:Protein Precursors; D017474:Receptors, Laminin / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D007797:Laminin; D011498:Protein Precursors; D017474:Receptors, Laminin