# Designing Functional Metalloproteins

> **NIH NIH R01** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2022 · $299,668

## Abstract

Metalloproteins are critical cellular workhorses, catalyzing difficult but essential transformations that are required
for functions as diverse as energy generation, metabolism, or signaling. The interplay between protein structure
and catalytic activity of these enzymes forms the foundation of our work. The protein may influence both the first
coordination sphere (ligand type, number, and relative geometry) as well as the outer coordination environment.
Despite the indispensable nature of this chemistry, a fundamental understanding of the relationship between
metal ion catalysts and a protein scaffold remains elusive. While native metalloenzyme studies provide valuable
insights, interpretations of these systems are often inadequate because of the intricacy of the scaffold or the
presence of multiple metal centers. Similarly, small molecules can afford insight on reactions but are often limited
by the solvent or synthetic complexity. For this reason, we utilize an intermediate approach with de novo design
of -helical proteins which are easily synthesized, water soluble, recapitulate native metalloprotein active sites
well, and provide native-like folds that can be manipulated simply in order to limit the variables that are probed
during our studies. Our long-term goal is to describe to full chemical resolution the processes by which metals
interact with three stranded coiled-coils (3SCC) composed of -helical proteins in order to achieve specific metal
structures and afford the desired catalysis. The overall objective is to insert metals into well-defined scaffolds
that can then be altered to test important hypotheses that will explain how these catalytic centers work. Our
central hypothesis is that well-defined de novo metalloproteins can be used to interrogate metal behavior within
designed proteins in ways that would be difficult to achieve with native proteins or small molecules. Our basic
premise is that de novo protein design provides simple, highly-controllable, and easily modified scaffolds that
are well suited to extract fundamental information on metal chemistry. These structures provide a framework to
examine and compare systematic perturbations to different coordination sites in aqueous peptidic environments,
which is often difficult to do with small molecule models, especially those insoluble in water. The rationale of the
proposed research is that it will provide new information on protein-metal interactions that have eluded the
scrutiny of other approaches due to the complexity or insolubility of the natural systems. These studies will eventually
form the basis for developing new catalysts with different functions using essentially the same protein ligands but differentmetals.
We can also now explore the influence of asymmetric first and second coordination sphere environments and changing the
helical twist on the structural and catalytic behavior of proteins. Our hypothesis will be tested through three Specific Aims:
1) Evaluatio...

## Key facts

- **NIH application ID:** 10374893
- **Project number:** 5R01GM141086-02
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** VINCENT L PECORARO
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $299,668
- **Award type:** 5
- **Project period:** 2021-04-01 → 2025-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10374893

## Citation

> US National Institutes of Health, RePORTER application 10374893, Designing Functional Metalloproteins (5R01GM141086-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10374893. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*