Identification of Setaria cervi heat shock protein 70 by mass spectrometry and its evaluation as diagnostic marker for lymphatic filariasis.
Using mass spectrometry and immunological approaches, a heat shock protein 70 associated with lymphatic filariasis (LF) has been identified from a bovine filarial parasite Setaria cervi. A heat shock protein was detected in different life stages of S. cervi when exposed to an elevated temperature of 44 degrees C. A combination of ATP-agarose column chromatography and electro-elution was used for its purification from adult female extract. On closer examination, it migrated as a single band at 68 kDa on 10% SDS-PAGE. Peptide sequences TTPSYVAFTDTER, DSGAIAGLNVLR, IINEPTAAAIAYGLDK, NALESYAFNMK and LLSDFFSGK were obtained through MALDI-LC/MS analysis. Confirmation of peptides was accomplished by MASCOT database which showed substantial sequence homology with S. digitata, Wuchereria bancrofti, and Caenorhabditis elegans. Multiple sequence alignment using Clustal W showed 98% identity with W. bancrofti and only 28% with human HSP70. Furthermore, the antigenicity plot has shown that the highly antigenic amino acid residues are constituted within the conserved peptides. These observations suggest a plausible biological connection of ScHSP70 with the disease and its strong immunogenic nature. ScHSP70 showed antigenic cross-reactivity with IgG class of antibody in different categories of filarial sera. However, when IgG subclasses were tested, IgG4 showed high specificity and sensitivity with asymptomatic microfilaraemic sera.
Srivastava
Saurabha
S
Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi-221005, India.
Srikanth
Elesela
E
Liebau
Eva
E
Rathaur
Sushma
S
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D015415:Biomarkers; D002417:Cattle; D003429:Cross Reactions / Q000235:genetics / Q000276:immunology; D004605:Elephantiasis, Filarial / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology*; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000276:immunology*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008297:Male; D013058:Mass Spectrometry; D010455:Peptides / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D017182:Setaria Nematode / Q000235:genetics / Q000276:immunology*; D014958:Wuchereria bancrofti / Q000235:genetics / Q000276:immunology*
D000909:Antibodies, Helminth; D015415:Biomarkers; D018840:HSP70 Heat-Shock Proteins; D015801:Helminth Proteins; D007074:Immunoglobulin G; D010455:Peptides