# De Novo Design of Functional Metalloporphyrin-Containing Proteins

> **NIH NIH F32** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 2020 · $65,310

## Abstract

Project Summary/Abstract
Metalloproteins perform chemical transformations with rates and selectivites that have yet to be achieved in
synthetic or designed systems. These differences in reactivity are directly linked to the environment produced
by the protein matrix that cannot be easily reproduced in synthetic constructs. To test our understanding of
how metalloproteins function, we aim to design de novo metalloproteins from scratch. Proteins that bind
porphyrin-like cofactors are of particular interest, as heme proteins are known to perform a variety of
reactions. Only recently have we been able to design proteins that bind a cofactor with sub-Å accuracy. This
opens the door to expand on the utility of natural proteins by incorporating synthetic inorganic cofactors into
proteins that lack pre-evolved function. The proposed research strategy seeks to elucidate the design features
necessary to bind M-tetraphenylporphyrin (M-TPP; M=Fe, Mn) complexes in close proximity to a substrate
binding pocket. First- and second-shell interactions will be engineered to control orientation, electronic
structure, and reaction pathway of the cofactor and substrate. Binding pockets will be designed for camphor
and styrene and these de novo metalloproteins will be tested for their activity towards hydroxylation and
epoxidation. The selectivity of these metalloenzymes will also be tested and the protein scaffold will be
redesigned to elicit regioselective reactions (i.e. reacting with only one of two possible functionalizable groups).
The proteins will be characterized using optical spectroscopies, electron paramagnetic resonance (EPR)
spectroscopy, NMR spectroscopy, and by X-ray diffraction (XRD) methods. This work would be a breakthrough
in protein design and will directly impact the fundamental understanding of the effects of protein
environments on the function of metal centers in metalloproteins.

## Key facts

- **NIH application ID:** 9850083
- **Project number:** 5F32GM130029-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **Principal Investigator:** Samuel I Mann
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $65,310
- **Award type:** 5
- **Project period:** 2018-12-01 → 2021-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9850083

## Citation

> US National Institutes of Health, RePORTER application 9850083, De Novo Design of Functional Metalloporphyrin-Containing Proteins (5F32GM130029-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9850083. Licensed CC0.

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