title: Characterization of a Cryptosporidium parvum gene encoding a protein with homology to long chain fatty acid synthetase.
abstract: We describe here the cloning, sequencing, and characterization of a novel Cryptosporidium parvum gene, encoding a protein with significant homology to the long-chain fatty acyl-CoA synthetase (LCFA, EC 6.2.1.3). The gene has an open reading frame of 2,301 bp, coding for a 766 amino acid polypeptide, and with an estimated MW of 86.1 kDa. By indirect immunofluorescence assay, monoclonal antibodies C3CE7 and E5D labeled the anterior pole of fixed C. parvum sporozoites and developmental stages in C. parvum-infected cultures at 24, 48, and 72 h post-infection. These monoclonal antibodies inhibited more than 35% of parasite growth in vitro. The effect of triacsin C, a potent selective inhibitor of LCFA synthetase, on parasite growth was assessed in cell culture; complete inhibition of parasite growth at 2.5 ug/ml was obtained with little evidence of drug-associated cytotoxicity. These results suggest that the fatty acyl-CoA synthetase may be a useful target in the development of selective inhibitors and immunologic interventions against C. parvum.
authors.affiliation: Centers for Disease Control and Prevention, Chamblee, GA 30341, USA.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Camero
authors.forename: William P
authors.initials: WP
authors.lastname: Shulaw
authors.forename: Lihua
authors.initials: L
authors.lastname: Xiao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics; D001665:Binding Sites; D002460:Cell Line; D003066:Coenzyme A Ligases / Q000235:genetics*; D016785:Cryptosporidium parvum / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000235:genetics*; D004285:Dogs; D015723:Gene Library; D017125:Genes, Protozoan; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D016053:RNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D012333:RNA, Messenger; D016053:RNA, Protozoan; D003066:Coenzyme A Ligases