Localization, turnover and conservation of gp15/400 in different stages of Brugia malayi.
The expression of a protein complex designated gp15/400, previously identified via extrinsic iodination of adult Brugia malayi, was examined by labelling all stages found in the mammalian host and immunoprecipitation with a specific antibody raised to a recombinant protein. In this way, gp15/400 could be detected in L3, L4, adult worms and microfilariae recovered from jirds and labelled with Bolton-Hunter reagent. Metabolic labelling indicated that gp15/400 was released into culture medium when adult worms were maintained in vitro, but at a rate slower than that of gp29, the major soluble cuticular glycoprotein. Immuno-electron microscopy showed that the protein complex was broadly distributed in different tissues, although it was not detectable in the cuticle of adult worms. Dense labelling was observed in the matrix of the basal laminae bordering the hypodermis, somatic musculature and oesophagus, and lower but significant labelling was seen in the cells overlying these extracellular matrices. Hybridization of genomic DNA with a cDNA probe encoding gp15/400 indicated that homologous genes were present in Dirofilaria immitis and Acanthocheilonema viteae. The failure to detect related genes in non-filarial nematodes was presumed to be due to divergence beyond the practical limits of detection by nucleic acid probes, as antibody reagents showed that the protein cross-reacted immunologically with ABA-1, a major protein allergen from the body fluid of Ascaris.
Selkirk
M E
ME
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London.
Gregory
W F
WF
Jenkins
R E
RE
Maizels
R M
RM
D000330:Aedes / Q000469:parasitology; D000818:Animals; D015139:Blotting, Southern; D017178:Brugia malayi / Q000378:metabolism*; D003429:Cross Reactions; D004247:DNA / Q000302:isolation & purification; D005260:Female; D017238:Genes, Helminth; D005849:Gerbillinae / Q000469:parasitology; D015801:Helminth Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D008297:Male; D016253:Microscopy, Immunoelectron; D009024:Morphogenesis / Q000502:physiology; D009348:Nematoda / Q000235:genetics / Q000276:immunology; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D015801:Helminth Proteins; D004247:DNA