Our program is devoted to answering key questions in biology through the development and application of multinuclear electron-nuclear double resonance (ENDOR) and electron spin-echo envelope modulation (ESEEM) spectroscopies to characterize transition-metal centers vital to human health and disease. Application of these techniques to intermediate states that are trapped by freeze-quench and cryoreduction methods, and are isotopically labeled by outstanding teams of collaborating chemists and biologists, can completely characterize the active-site environment of a metal ion at every stage of an enzyme's catalytic cycle. As augmented by novel kinetic protocols we have developed, these studies reveal in precise details the function of critical metal centers, not only in isolate enzymes but in living cells as well. Dramatic progress during our current grant period has inspired multiple specific aims: (i) Small-molecule activation by metalloenzymes, including Dioxygen-activation by Heme Monooxygenases and Dinitrogen Reduction by Nitrogenase; (ii) Biomimetic Mo and Fe Complexes as Models for Nitrogenase Intermediates and Paradigms for the Jahn-Taller Effect in trigonal symmetry; (iii) Radical Reactions in Metalloenzymes -The Radical SAM Superfamily; (iv) in vivo Speciation of Mn(II) - Protection from Oxidative Stress, Mechanisms of Toxicity, Probe of Physiology; (v) ATPase Co(II)/Mn(II) Transporters; (vi) ENDOR Methodology Development - Hyperfine Signs, Electron-Nuclear-Electron Triple Resonance, Q-band Resonator Development; (vii) Extending the Biological Applications of ENDOR Techniques. Many of these aims reflect longstanding efforts, while projects associated with transition-metal homeostasis that emerged this period (Mn(II) speciation, and Mn(II)/Co(II) transport) reflect a commitment to opening new areas, as explicitly expressed in the final Aim.

Public Health Relevance

Metal centers play a central role in human health and disease. Our program is devoted to the development and application of advanced paramagnetic resonance measurement techniques (ENDOR/ESEEM) as a uniquely incisive means of revealing how biologically important metal centers function. The systems to be studied play many essential roles, including: protection from oxidative stress and reactive-oxygen species (ROS); formation of the key signaling and immune-response molecules, nitric oxide and carbon monoxide; xenobiotic and drug metabolism; formation of essential cofactors, whose absence through genetic deficiencies is deadly; homeostasis of metal ions (Mn/Co/Fe) whose regulation in humans and pathogens is central in health and disease; enzymatic formation of the bio-available nitrogen on which two-thirds of the world's population depend.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM111097-45S1
Application #
9608905
Study Section
Program Officer
Anderson, Vernon
Project Start
1979-01-01
Project End
2018-07-31
Budget Start
2017-02-01
Budget End
2018-07-31
Support Year
45
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Northwestern University at Chicago
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
160079455
City
Evanston
State
IL
Country
United States
Zip Code
60201
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