Identification of a neutrophil chemotactic factor from Tritrichomonas foetus as superoxide dismutase.
Antibodies to a neutrophil chemotactic factor from Tritrichomonas foetus were used to screen a T. foetus cDNA expression library in lambda gt11. All positive clones were identified as homologs of iron-containing superoxide dismutase (SOD). Native gel electrophoresis showed that the antibodies indeed recognized T. foetus antigens with SOD activity. Two SOD genes were found in T. foetus, and cloned and sequenced as parts of larger genomic segments of 3844 and 4089 base pairs. Transcription initiated between the first and second methionine codons of each genomic open reading frame, generating mRNAs with 5' untranslated regions of 11-15 bases, and encoding proteins of 195 amino acids. The two SOD coding sequences lacked obvious introns. They were 79% identical at both the nucleotide and amino acid levels. Both SOD genes were inserted into a eukaryotic expression vector and stably expressed in mammalian cells; both proteins were recognized by the antibodies, and both assumed a cytosolic, extranuclear distribution in these cells. Histidine-tagged forms of both T. foetus SODs were expressed in E. coli and after purification, found to have neutrophil chemotactic activity similar to the non-recombinant factor purified from T. foetus. Identification of this neutrophil chemotactic factor as SOD provides additional insight into the host-parasite interaction.
Granger
B L
BL
Veterinary Molecular Biology Laboratory, Montana State University, Bozeman 59717-3610, USA. uvsbg@montana.edu
Warwood
S J
SJ
Hayai
N
N
Hayashi
H
H
Owhashi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D006224:Cricetinae; D016209:Interleukin-8 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000276:immunology / Q000469:parasitology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D016835:Tritrichomonas foetus / Q000201:enzymology* / Q000235:genetics
D016209:Interleukin-8; D013482:Superoxide Dismutase