Robust and Versatile Host Protein for the Design and Evaluation of Artificial Metal Centers
Type
ArticleAuthors
Fischer, Johannes
Renn, Dominik
Quitterer, Felix
Radhakrishnan, Anand
Liu, Meina
Makki, Arwa

Ghoprade, Seema Arun

Rueping, Magnus

Arold, Stefan T.

Groll, Michael

Eppinger, Jörg

KAUST Department
Biological & Organometallic Catalysis LaboratoriesBiological and Environmental Sciences and Engineering (BESE) Division
Bioscience Program
Chemical Engineering Program
Chemical Science Program
Computational Bioscience Research Center (CBRC)
KAUST Catalysis Center (KCC)
KAUST Catalysis Center (KCC), Division of Physical Sciences & Engineering and § Computational Bioscience Research Center (CBRC), Division of Biological Sciences & Engineering, King Abdullah University of Science and Technology (KAUST), 23955 Thuwal, MK, Saudi Arabia
Office of the VP
Physical Science and Engineering (PSE) Division
Structural Biology and Engineering
Date
2019-11-12Embargo End Date
2020-11-12Permanent link to this record
http://hdl.handle.net/10754/660353
Metadata
Show full item recordAbstract
Artificial metalloenzymes (ArMs) have high potential in biotechnological applications as they combine the versatility of transition-metal catalysis with the substrate selectivity of enzymes. An ideal host protein should allow high-yield recombinant expression, display thermal and solvent stability to withstand harsh reaction conditions, lack nonspecific metal-binding residues, and contain a suitable cavity to accommodate the artificial metal site. Moreover, to allow its rational functionalization, the host should provide an intrinsic reporter for metal binding and structural changes, which should be readily amendable to high-resolution structural characterization. Herein, we present the design, characterization, and de novo functionalization of a fluorescent ArM scaffold, named mTFP*, that achieves these characteristics. Fluorescence measurements allowed direct assessment of the scaffold's structural integrity. Protein X-ray structures and transition metal Förster resonance energy transfer (tmFRET) studies validated the engineered metal coordination sites and provided insights into metal binding dynamics at the atomic level. The implemented active metal centers resulted in ArMs with efficient Diels-Alderase and Friedel-Crafts alkylase activities.Citation
Fischer, J., Renn, D., Quitterer, F., Radhakrishnan, A., Liu, M., Makki, A., … Eppinger, J. (2019). Robust and Versatile Host Protein for the Design and Evaluation of Artificial Metal Centers. ACS Catalysis, 11371–11380. doi:10.1021/acscatal.9b02896Sponsors
The staff of the Beamline X06SA at the Paul Scherrer Institute, SLS, Villigen, Switzerland, is acknowledged for assistance during data collection. The authors thank S. Abdul Rajjaka for help with the bioinformatic setup.Publisher
American Chemical Society (ACS)Journal
ACS CatalysisAdditional Links
https://pubs.acs.org/doi/10.1021/acscatal.9b02896ae974a485f413a2113503eed53cd6c53
10.1021/acscatal.9b02896