Show simple item record

dc.contributor.authorFischer, Johannes
dc.contributor.authorRenn, Dominik
dc.contributor.authorQuitterer, Felix
dc.contributor.authorRadhakrishnan, Anand
dc.contributor.authorLiu, Meina
dc.contributor.authorMakki, Arwa
dc.contributor.authorGhoprade, Seema Arun
dc.contributor.authorRueping, Magnus
dc.contributor.authorArold, Stefan T.
dc.contributor.authorGroll, Michael
dc.contributor.authorEppinger, Jörg
dc.date.accessioned2019-12-02T08:52:11Z
dc.date.available2019-12-02T08:52:11Z
dc.date.issued2019-11-12
dc.identifier.citationFischer, 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.9b02896
dc.identifier.doi10.1021/acscatal.9b02896
dc.identifier.urihttp://hdl.handle.net/10754/660353
dc.description.abstractArtificial 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.
dc.description.sponsorshipThe 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.
dc.publisherAmerican Chemical Society (ACS)
dc.relation.urlhttps://pubs.acs.org/doi/10.1021/acscatal.9b02896
dc.titleRobust and Versatile Host Protein for the Design and Evaluation of Artificial Metal Centers
dc.typeArticle
dc.contributor.departmentBiological & Organometallic Catalysis Laboratories
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentChemical Engineering Program
dc.contributor.departmentChemical Science Program
dc.contributor.departmentComputational Bioscience Research Center (CBRC)
dc.contributor.departmentKAUST Catalysis Center (KCC)
dc.contributor.departmentKAUST 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
dc.contributor.departmentOffice of the VP
dc.contributor.departmentPhysical Science and Engineering (PSE) Division
dc.contributor.departmentStructural Biology and Engineering
dc.identifier.journalACS Catalysis
dc.rights.embargodate2020-11-12
dc.eprint.versionPost-print
dc.contributor.institutionCenter for Integrated Protein Science, Department Chemie, Lehrstuhl für Biochemie, Technische Universität München (TUM), D-85747 Garching, Germany
dc.contributor.institutionCentre de Biochimie Structurale, CNRS, INSERM, Université de Montpellier, 34090 Montpellier, France
kaust.personFischer, Johannes
kaust.personRenn, Dominik
kaust.personRadhakrishnan, Anand
kaust.personLiu, Meina
kaust.personMakki, Arwa
kaust.personGhorpade, Seema
kaust.personRueping, Magnus
kaust.personArold, Stefan T.
kaust.personEppinger, Jorg


This item appears in the following Collection(s)

Show simple item record