Molecular LEGO by domain-imprinting of cytochrome P450 BM3

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dc.contributor.authorJetzschmann, Katharina J.
dc.contributor.authorYarman, Aysu
dc.contributor.authorRustam, L.
dc.contributor.authorKielb, P.
dc.contributor.authorUrlacher, V. B.
dc.contributor.authorFischer, A.
dc.contributor.authorScheller, Frieder W.
dc.date.accessioned2021-01-08T21:51:24Z
dc.date.available2021-01-08T21:51:24Z
dc.date.issued2018
dc.departmentTAÜ, Fen Fakültesi, Moleküler Biyoteknoloji Bölümüen_US
dc.descriptionWeidinger, Inez/0000-0001-9316-6349; Fischer, Anna T. L./0000-0003-4567-3009en_US
dc.descriptionWOS:000429762100028en_US
dc.descriptionPubMed: 29413602en_US
dc.description.abstractHypothesis: Electrosynthesis of the MIP nano-film after binding of the separated domains or holocytochrome BM3 via an engineered anchor should result in domain-specific cavities in the polymer layer. Experiments: Both the two domains and the holo P450 BM3 have been bound prior polymer deposition via a N-terminal engineered his6-anchor to the electrode surface. Each step of MIP preparation was characterized by cyclic voltammetry of the redox-marker ferricyanide. Rebinding after template removal was evaluated by quantifying the suppression of the diffusive permeability of the signal for ferricyanide and by the NADH-dependent reduction of cytochrome c by the reductase domain (BMR). Findings: The working hypothesis is verified by the discrimination of the two domains by the respective MIPs: The holoenzyme P450 BM3 was ca. 5.5 times more effectively recognized by the film imprinted with the oxidase domain (BMO) as compared to the BMR-MIP or the non-imprinted polymer (NIP). Obviously, a cavity is formed during the imprinting process around the hiss-tag-anchored BMR which cannot accommodate the broader BMO or the P450 BM3. The affinity of the MIP towards P450 BM3 is comparable with that to the monomer in solution. The hiss-tagged P450 BM3 binds (30 percent) stronger which shows the additive effect of the interaction with the MIP and the binding to the electrode. (C) 2018 Published by Elsevier B.V.
dc.description.sponsorshipBMBF of GermanyFederal Ministry of Education & Research (BMBF) [0311993]; ERA-Chemistry [61133, OKTA NN117637]; Deutsche Forschungsgemeinschaft (DFG)German Research Foundation (DFG) [EXC 314]; Deutsche Forschungsgemeinschaft (DFG) and its graduate school (Berlin International Graduate School of Natural Sciences and Engineering); Freiburg University; BMBFFederal Ministry of Education & Research (BMBF) [FKZ: 01FP13033F]
dc.description.sponsorshipThe authors would like to thank BMBF (0311993) of Germany well as ERA-Chemistry (2014, 61133; OKTA NN117637) for the financial support. This work is a part of UniCat, the Cluster of Excellence in the field of catalysis coordinated by the Technical University of Berlin and financially supported by Deutsche Forschungsgemeinschaft (DFG) within the framework of the German Excellence Initiative (EXC 314) and its graduate school (Berlin International Graduate School of Natural Sciences and Engineering). In addition, AF would like to thank financial support from Freiburg University as well as from the BMBF (FKZ: 01FP13033F).
dc.identifier.doi10.1016/j.colsurfb.2018.01.047
dc.identifier.endpage246en_US
dc.identifier.issn0927-7765
dc.identifier.issn1873-4367
dc.identifier.scopus2-s2.0-85041461334
dc.identifier.scopusqualityQ1
dc.identifier.startpage240en_US
dc.identifier.urihttp://doi.org/10.1016/j.colsurfb.2018.01.047
dc.identifier.urihttps://hdl.handle.net/20.500.12846/225
dc.identifier.volume164en_US
dc.identifier.wosWOS:000429762100028
dc.identifier.wosqualityQ1
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.institutionauthorYarman, Aysu
dc.language.isoen
dc.publisherElsevier Science Bv
dc.relation.ispartofColloids And Surfaces B-Biointerfaces
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectMolecularly Imprinted Polymersen_US
dc.subjectProtein Imprintingen_US
dc.subjectElectropolymerizationen_US
dc.subjectCytochrome P450en_US
dc.titleMolecular LEGO by domain-imprinting of cytochrome P450 BM3
dc.typeArticle

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