Characterization of magnesium requirement of human 5'-tyrosyl DNA phosphodiesterase mediated reaction

被引：11

作者：

Sanjay Adhikari

Soumendra K Karmahapatra

Tejaswita M Karve

Sanjona Bandyopadhyay

Jordan Woodrick

Praveen V Manthena

Eric Glasgow

Stephen Byers

Tapas Saha

Aykut Uren

机构：

[1] Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington

[2] Department of Biochemistry and Molecular and Cellular Biology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington

来源：

BMC Research Notes | / 5卷 / 1期

基金：

美国国家卫生研究院;

关键词：

Product Formation; Cold Ringer; Phosphotyrosyl; Excess EDTA; Active Site Tyrosine;

D O I：

10.1186/1756-0500-5-134

中图分类号：

学科分类号：

摘要：

Background: Topo-poisons can produce an enzyme-DNA complex linked by a 3'- or 5'-phosphotyrosyl covalent bond. 3'-phosphotyrosyl bonds can be repaired by tyrosyl DNA phosphodiesterase-1 (TDP1), an enzyme known for years, but a complementary human enzyme 5'-tyrosyl DNA phosphodiesterase (hTDP2) that cleaves 5'-phosphotyrosyl bonds has been reported only recently. Although hTDP2 possesses both 3'- and 5'- tyrosyl DNA phosphodiesterase activity, the role of Mg 2+ in its activity was not studied in sufficient details. Results: In this study we showed that purified hTDP2 does not exhibit any 5'-phosphotyrosyl phosphodiesterase activity in the absence of Mg 2+/Mn 2+, and that neither Zn 2+ or nor Ca 2+ can activate hTDP2. Mg 2+ also controls 3'-phosphotyrosyl activity of TDP2. In MCF-7 cell extracts and de-yolked zebrafish embryo extracts, Mg 2+ controlled 5'-phosphotyrosyl activity. This study also showed that there is an optimal Mg 2+ concentration above which it is inhibitory for hTDP2 activity. Conclusion: These results altogether reveal the optimal Mg 2+ requirement in hTDP2 mediated reaction. © 2011 Adhikari et al; licensee BioMed Central Ltd.

引用

共 28 条

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