Cloning and characterization of glutamate dehydrogenase (GDH) from the gut of Haemonchus contortus.
Vaccination of lambs with the membrane-bound (S3) thiol-Sepharose binding protein (TSBP) fraction derived from the gut of Haemonchus contortus confers significant protection against homologous challenge. The S3 TSBP peptide profile is dominated by a major protein of ca. 60 kDa which is strongly recognized by antisera from sheep demonstrably protected following immunization with S3 TSBP. In an attempt to identify this protein, sera from protected lambs were employed to screen a lambda gt11 cDNA library of the adult parasite and resulted in the isolation of numerous clones encoding a homologue of the mitochondrial enzyme, glutamate dehydrogenase (GDH). GDH enzyme activity was readily demonstrable in S3 TSBP material and immunolocalization studies showed that the enzyme was localized to the cytoplasm of the parasite's gut. Furthermore, the enzyme appeared to be developmentally regulated, with both GDH mRNA and protein expressed almost exclusively during the blood-feeding parasitic stages.
Skuce
P J
PJ
Moredun Research Institute, International Research Centre, Penicuik, Midlothian, UK.
Stewart
E M
EM
Smith
W D
WD
Knox
D P
DP
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000032:analysis; D029968:Escherichia coli Proteins; D018507:Gene Expression Regulation, Developmental; D005969:Glutamate Dehydrogenase / Q000235:genetics* / Q000378:metabolism*; D006188:Haemonchiasis / Q000469:parasitology; D006189:Haemonchus / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D033902:Periplasmic Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014611:Vaccination
D000947:Antigens, Helminth; D002352:Carrier Proteins; C094586:CysP protein, E coli; D018509:DNA, Helminth; D029968:Escherichia coli Proteins; D033902:Periplasmic Binding Proteins; D005969:Glutamate Dehydrogenase