A novel nuclease activity that is activated by Ca(2+) chelated to EGTA.

Systems Biology in Reproductive Medicine
Kenneth Dominguez, W Steven Ward

Abstract

Most nucleases require a divalent cation as a cofactor, usually Mg(2+) or Ca(2+), and are inhibited by the chelators EDTA and EGTA. We report the existence of a novel nuclease activity, initially identified in the luminal fluids of the mouse male reproductive tract but subsequently found in other tissues,that requires EGTA chelated to calcium to digest DNA. We refer to this unique enzyme as CEAN (Chelated EGTA Activated Nuclease). Using a fraction of vas deferens luminal fluid, plasmid DNA was degraded in the presence of excess Ca(2+) (Ca(2+) :EGTA = 16) or excess EGTA (Ca(2+) :EGTA = 0.25), but required the presence of both. Higher levels of EGTA (Ca(2+) :EGTA = 0.10) prevented activity, suggesting that unchelated EGTA may be a competitive inhibitor. The EGTA-Ca(2+) activation of CEAN is reversible as removing EGTA-Ca(2+) stops ongoing DNA degradation, but adding EGTA-Ca(2+) again reactivates the enzyme. This suggests the possibility that CEAN binds directly to EGTA-Ca(2+). CEAN has a greater specificity for the chelator than for the divalent cation. Two other chelators, BAPTA and sodium citrate, do not activate CEAN in the presence of cation, but chelated EDTA does. EGTA chelated to other divalent cations such as Mn(2+), Zn(2...Continue Reading

References

Jan 1, 1984·The American Journal of Physiology·D J Miller, G L Smith
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Dec 4, 2003·Gene·Cory J Evans, Renato J Aguilera
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Citations

Dec 30, 2010·The Journal of Biological Chemistry·Aicha Jrad-LamineJoël R Drevet
Nov 14, 2013·PloS One·Alan M ElderCandace K Mathiason
Aug 1, 2012·Reproduction in Domestic Animals = Zuchthygiene·R J AitkenM A Baker
Oct 16, 2015·Human Reproduction·Joanna E GaweckaW Steven Ward
Mar 25, 2019·Biotechnology and Applied Biochemistry·Yuxiang LiuZeying Liu
Jan 6, 2011·Systems Biology in Reproductive Medicine·Michael A OrtegaW Steven Ward
Oct 3, 2018·Frontiers in Endocrinology·Donatella PaoliFrancesco Lombardo
Feb 23, 2020·Applied Microbiology and Biotechnology·Lynn Sophie SchwardmannSkander Elleuche

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