Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism.

Lipton, Andrew S; Heck, Robert W; Ellis, Paul D

doi:10.1021/ja0305609

Title: Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism.

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Abstract

Many zinc enzymes utilize zinc bound water as a critical component of a catalytic reaction. The Zn2+ ion activates water through ionization, polarization, or simple displacement depending upon the mechanistic details. The fate of one proton from the bound water is determined primarily by the influence of directly bound Zn-ligands, as well as hydrogen bonding with a secondary coordination sphere of side chains and/or bound waters within the protein. We have employed low temperature solid-state 67Zn NMR spectroscopy to probe the nature of the bonding at Zn2+ in human carbonic anhydrase isozyme II (CAII). In particular we wanted to characterize the 67Zn NMR parameters of the metal with both water and hydroxide as the fourth ligand, but instead we show that hydroxide is bound to Zn2+ over the pH range of 5 to 8.5. These results suggest the accepted mechanism of action of CAII needs to be revised. These data serve to provide further understanding of the observed pH dependence of the activity of this well studied protein.

Authors:: Lipton, Andrew S; Heck, Robert W; Ellis, Paul D

Publication Date:: Wed Apr 14 00:00:00 EDT 2004

Research Org.:: Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

Sponsoring Org.:: USDOE

OSTI Identifier:: 15007489

Report Number(s):: PNWD-SA-6180

DOE Contract Number:: AC06-76RL01830

Resource Type:: Journal Article

Journal Name:: Journal of the American Chemical Society, 126(14):4735-4739

Additional Journal Information:: Journal Name: Journal of the American Chemical Society, 126(14):4735-4739

Country of Publication:: United States

Language:: English

Subject:: 08 HYDROGEN

Citation Formats


                    Lipton, Andrew S, Heck, Robert W, and Ellis, Paul D. Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism..  United States: N. p., 2004. 
        Web.  doi:10.1021/ja0305609.

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                    Lipton, Andrew S, Heck, Robert W, & Ellis, Paul D. Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism..  United States.  https://doi.org/10.1021/ja0305609

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                    Lipton, Andrew S, Heck, Robert W, and Ellis, Paul D. 2004.  
        "Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism.".  United States.  https://doi.org/10.1021/ja0305609.

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@article{osti_15007489,

  title        = {Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism.},

  author       = {Lipton, Andrew S and Heck, Robert W and Ellis, Paul D},

  abstractNote = {Many zinc enzymes utilize zinc bound water as a critical component of a catalytic reaction. The Zn2+ ion activates water through ionization, polarization, or simple displacement depending upon the mechanistic details. The fate of one proton from the bound water is determined primarily by the influence of directly bound Zn-ligands, as well as hydrogen bonding with a secondary coordination sphere of side chains and/or bound waters within the protein. We have employed low temperature solid-state 67Zn NMR spectroscopy to probe the nature of the bonding at Zn2+ in human carbonic anhydrase isozyme II (CAII). In particular we wanted to characterize the 67Zn NMR parameters of the metal with both water and hydroxide as the fourth ligand, but instead we show that hydroxide is bound to Zn2+ over the pH range of 5 to 8.5. These results suggest the accepted mechanism of action of CAII needs to be revised. These data serve to provide further understanding of the observed pH dependence of the activity of this well studied protein.},

  doi          = {10.1021/ja0305609},

  url          = {https://www.osti.gov/biblio/15007489},
  journal      = {Journal of the American Chemical Society, 126(14):4735-4739},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {Wed Apr 14 00:00:00 EDT 2004},

  month        = {Wed Apr 14 00:00:00 EDT 2004}

}

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