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Human eosinophil-derived neurotoxin: involvement of a putative non-catalytic phosphate-binding subsite in its catalysis

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Abstract

Human eosinophil-derived neurotoxin (EDN) or RNase 2, found in the non-core matrix of eosinophils is a ribonuclease belonging to the Ribonuclease A superfamily. EDN manifests a number of bioactions including neurotoxic and antiviral activities, which are dependent on its ribonuclease activity. The core of the catalytic site of EDN contains various base and phosphate-binding subsites. Unlike many members of the RNase A superfamily, EDN contains an additional non-catalytic phosphate-binding subsite, P−1. Although RNase A also contains a P−1 subsite, the composition of the site in EDN and RNase A is different. In the current study we have generated site-specific mutants to study the role of P−1 subsite residues Arg36, Asn39, and Gln40 of EDN in its catalytic activity. The individual mutation of Arg36, Asn 39, and Gln40 resulted in a reduction in the catalytic activity of EDN on poly(U) and poly(C). However, there was no change in the activities on yeast tRNA and dinucleotide substrates. The study shows that the P−1 subsite is crucial for the ribonucleolytic activity of EDN on polymeric RNA substrates.

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Acknowledgments

This work was supported by grants to the National Institute of Immunology from the Department of Biotechnology, Government of India, and a grant (SP/SO/BB-01/2003) to JKB from the Department of Science and Technology, Government of India. DS, DS, and PD are Senior Research Fellows of the Council of Scientific and Industrial Research, India.

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Authors and Affiliations

  1. Immunochemistry Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Deepa Sikriwal, Divya Seth, Punyatirtha Dey & Janendra K. Batra

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  1. Deepa Sikriwal
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  2. Divya Seth
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  3. Punyatirtha Dey
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  4. Janendra K. Batra
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Corresponding author

Correspondence to Janendra K. Batra.

Additional information

Deepa Sikriwal and Divya Seth contributed equally to this work.

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Sikriwal, D., Seth, D., Dey, P. et al. Human eosinophil-derived neurotoxin: involvement of a putative non-catalytic phosphate-binding subsite in its catalysis. Mol Cell Biochem 303, 175–181 (2007). https://doi.org/10.1007/s11010-007-9471-0

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