Nitric Oxide Modulates Endonuclease III Redox Activity by a 800 mV Negative Shift upon [Fe4 S4 ] Cluster Nitrosylation

Journal of the American Chemical Society
Levi A EkangerJacqueline K Barton

Abstract

Here we characterize the [Fe4S4] cluster nitrosylation of a DNA repair enzyme, endonuclease III (EndoIII), using DNA-modified gold electrochemistry and protein film voltammetry, electrophoretic mobility shift assays, mass spectrometry of whole and trypsin-digested protein, and a variety of spectroscopies. Exposure of EndoIII to nitric oxide under anaerobic conditions transforms the [Fe4S4] cluster into a dinitrosyl iron complex, [(Cys)2Fe(NO)2]-, and Roussin's red ester, [(μ-Cys)2Fe2(NO)4], in a 1:1 ratio with an average retention of 3.05 ± 0.01 Fe per nitrosylated cluster. The formation of the dinitrosyl iron complex is consistent with previous reports, but the Roussin's red ester is an unreported product of EndoIII nitrosylation. Hyperfine sublevel correlation (HYSCORE) pulse EPR spectroscopy detects two distinct classes of NO with 14N hyperfine couplings consistent with the dinitrosyl iron complex and reduced Roussin's red ester. Whole-protein mass spectrometry of EndoIII nitrosylated with 14NO and 15NO support the assignment of a protein-bound [(μ-Cys)2Fe2(NO)4] Roussin's red ester. The [Fe4S4]2+/3+ redox couple of DNA-bound EndoIII is observable using DNA-modified gold electrochemistry, but nitrosylated EndoIII does not di...Continue Reading

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Citations

Jun 22, 2019·Annual Review of Biochemistry·Jacqueline K BartonElizabeth O'Brien
Sep 10, 2019·Dalton Transactions : an International Journal of Inorganic Chemistry·Daniel B GrabarczykKylie A Vincent
May 30, 2020·Journal of the American Chemical Society·Rebekah M B SilvaJacqueline K Barton
Jul 30, 2020·Nitric Oxide : Biology and Chemistry·Peter C Ford, Katrina M Miranda
May 21, 2021·Inorganic Chemistry·Daniela R TruzziPeter C Ford
Aug 3, 2021·Chemical Communications : Chem Comm·D Chase PectolMarcetta Y Darensbourg
Sep 20, 2019·Inorganic Chemistry·Daniela R TruzziPeter C Ford
Sep 25, 2021·Journal of the American Chemical Society·Miguel N PintoJacqueline K Barton

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