Identification and characterization of Paragonimus westermani leucine aminopeptidase.
Paragonimus westermani is a tissue-invading trematode parasite that causes inflammatory lung disease as well as systemic infections including cerebral invasion in carnivorous mammals. While aminopeptidases play important roles in trematodes in the catabolism of host hemoglobin, an essential source of nutrient for the parasite, little is known about aminopeptidase in Paragonimus. Presently, we isolated a cDNA encoding a 58 kDa P. westermani leucine aminopeptidase (PwLAP). Deduced amino acid sequence of PwLAP exhibited significant sequence homology with LAP from Schistosoma spp. and Fasciola hepatica. Biochemical analysis of the recombinant PwLAP protein demonstrated preferential substrate specificity for Leu-NHMec and inhibition by EDTA, 1,10-phenanthroline, and bestatin, which are conserved characteristics of the M17 family of leucine aminopeptidase. PwLAP exhibited relatively higher enzyme activity in the presence of Mn2+ compared to Schistosoma mansoni LAP. Based on the biochemical properties and immunohistochemical analysis, PwLAP is concluded to represent a leucine aminopeptidase. The enzyme is most likely responsible for the catabolism of host hemoglobin, and, hence, represents a potential target of Paragonimus chemotherapy.
Song
Su-Min
SM
Department of Parasitology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
Park
Joon-Hyung
JH
Kim
Jin
J
Kim
Suk-Il
SI
Hong
Yeon-Chul
YC
Kong
Hyun-Hee
HH
Chung
Dong-Il
DI
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004064:Digestive System / Q000166:cytology* / Q000201:enzymology; D004283:Dog Diseases / Q000276:immunology / Q000469:parasitology / Q000473:pathology*; D004285:Dogs; D004848:Epithelium / Q000201:enzymology / Q000469:parasitology*; D005786:Gene Expression Regulation; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007931:Leucyl Aminopeptidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010237:Paragonimiasis / Q000276:immunology / Q000469:parasitology / Q000473:pathology / Q000662:veterinary; D048492:Paragonimus westermani / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D010802:Phylogeny; D051381:Rats; D017207:Rats, Sprague-Dawley; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D006454:Hemoglobins; D007931:Leucyl Aminopeptidase