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Setaria cervi dual specific phosphatase: characterization and its effect on eosinophil degranulation

Published online by Cambridge University Press:  15 June 2009

S. RATHAUR ,
R. RAI ,
E. SRIKANTH  and
S. SRIVASTAVA
S. RATHAUR*
Affiliation:
Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi-221005, India
R. RAI
Affiliation:
Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi-221005, India
E. SRIKANTH
Affiliation:
Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi-221005, India
S. SRIVASTAVA
Affiliation:
Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi-221005, India
*
*Corresponding author. Tel: +91 9336463404. E-mail: [email protected]
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Summary

Setaria cervi, a bovine filarial parasite contains significant acid phosphatase (AcP) activity in its various life stages. Two forms of AcP were separated from somatic extract of adult female parasite using cation exchange, gel filtration and concavalin affinity chromatography. One form having a molecular mass of 79 kDa was characterized as dual specific protein tyrosine phosphatase (ScDSP) based on substrate specificity and inhibition studies. With various substrates tested, it showed significant activity in the order of phospho-L-tyrosine>pNPP>ADP>phospho-L-serine. Inhibition by orthovanadate, fluoride, molybdate, and zinc ions further confirms protein tyrosine phosphatase nature of the enzyme. Km and Vmax determined with various substrates were found to be 16·66 mM, 25·0 μM/ml/min with pNPP; 20·0 mM, 40·0 μM/ml/min with phospho-L-tyrosine and 27·0 mM, 25·0 μM/ml/min with phospho-L-serine. KI with pNPP and sodium orthovanadate (IC50 33·0 μM) was calculated to be 50·0 mM. Inhibition with pHMB, silver nitrate, DEPC and EDAC suggested the presence of cysteine, histidine and carboxylate residues at its active site. Cross-reactivity with W. bancrofti-infected sera was demonstrated by Western blotting. ScDSP showed elevated levels of IgE in chronic filarial sera using ELISA. Under in vitro conditions, ScDSP resulted in increased effector function of human eosinophils when stimulated by IgG, which showed a further decrease with increasing enzyme concentration. Results presented here suggest that S. cervi DSP should be further studied to determine its role in pathogenesis and the persistence of filarial parasite.

Keywords

filariasisSetaria cerviacid phosphataseDSPIgEeosinophil degranulation
Type
Research Article
Information
Parasitology , Volume 136 , Issue 8 , July 2009 , pp. 895 - 904
Copyright
Copyright © Cambridge University Press 2009

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