Effects of mutating specific residues present near the amino terminus of 2'-5'-oligoadenylate synthetase.

The Journal of Biological Chemistry
A GhoshG C Sen

Abstract

In this study, we investigated the role of specific amino acid residues present near the amino terminus of the 9-2 isozyme of 2'-5'-oligoadenylate synthetase. In vitro expression of deletion mutants showed that residues 1-9 are required for enzyme activity. Within this region, residues 3, 7, and 8 were found to be conserved among all known isozymes of 2'-5'-oligoadenylate synthetase. Mutation of these residues singly or in combination resulted in partial or total loss of enzyme activity. Substitution of the proline residue at position 7 by different residues caused a partial or complete loss of activity. The properties of the inactive P7Q mutant were further explored by expressing the protein in bacteria. The bacterially expressed protein was also enzymatically inactive. The mutant protein could bind the substrate ATP and the activator double-stranded RNA normally. Oligomerization properties of the protein were examined by an affinity-based interaction assay and by glycerol gradient centrifugation; there was no detectable difference between the wild type and the P7Q mutant. These results demonstrated the importance of the proline residue at position 7 in conferring enzyme activity to the protein without affecting its other prop...Continue Reading

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Citations

Nov 12, 2009·Journal of Molecular Evolution·Karina Hansen KjaerJust Justesen
Feb 11, 2004·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Shoudong Li, Ganes C Sen
Feb 28, 2006·Journal of Virology·Svetlana V ScherbikMargo A Brinton
Jul 13, 2002·Proceedings of the National Academy of Sciences of the United States of America·Mee-Jung KimRobert M Krug
Jun 25, 2002·Proceedings of the National Academy of Sciences of the United States of America·Andrey A PerelyginMargo A Brinton
Mar 25, 2009·Expert Opinion on Drug Safety·Cristina Domingo, Matthias Niedrig
Aug 11, 2000·Seminars in Cancer Biology·G C Sen
Jan 6, 2018·Immune Network·Eunhye KimSoohyun Kim
Aug 28, 1999·The Journal of Biological Chemistry·S N SarkarG C Sen
Jan 9, 1999·The Journal of Biological Chemistry·S N SarkarG C Sen
Nov 17, 2016·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Hassan T Tag-El-Din-HassanTakashi Agui
Sep 15, 1998·Methods : a Companion to Methods in Enzymology·S N Sarkar, G C Sen
Nov 27, 2021·Science China. Life Sciences·Xiaoxue WangYinhua Huang

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