Redox Partner Interaction Sites in Cytochrome P450 Monooxygenases:In SilicoAnalysis and Experimental Validation

ChemistrySelectVolume 1, Issue 6 p. 1243-1251 Communication Redox Partner Interaction Sites in Cytochrome P450 Monooxygenases: In Silico Analysis and Experimental Validation Dr. Łukasz Gricman, Dr. Łukasz Gricman Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany These authors contributed equally to this work.Search for more papers by this authorDr. Martin J. Weissenborn, Dr. Martin J. Weissenborn Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany These authors contributed equally to this work.Search for more papers by this authorSara M. Hoffmann, Sara M. Hoffmann Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany These authors contributed equally to this work.Search for more papers by this authorNiels Borlinghaus, Niels Borlinghaus Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanySearch for more papers by this authorProf. Dr. Bernhard Hauer, Prof. Dr. Bernhard Hauer Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanySearch for more papers by this authorProf. Dr. Jürgen Pleiss, Corresponding Author Prof. Dr. Jürgen Pleiss Juergen.Pleiss@itb.uni-stuttgart.de Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanySearch for more papers by this author Dr. Łukasz Gricman, Dr. Łukasz Gricman Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany These authors contributed equally to this work.Search for more papers by this authorDr. Martin J. Weissenborn, Dr. Martin J. Weissenborn Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany These authors contributed equally to this work.Search for more papers by this authorSara M. Hoffmann, Sara M. Hoffmann Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany These authors contributed equally to this work.Search for more papers by this authorNiels Borlinghaus, Niels Borlinghaus Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanySearch for more papers by this authorProf. Dr. Bernhard Hauer, Prof. Dr. Bernhard Hauer Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanySearch for more papers by this authorProf. Dr. Jürgen Pleiss, Corresponding Author Prof. Dr. Jürgen Pleiss Juergen.Pleiss@itb.uni-stuttgart.de Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanySearch for more papers by this author First published: 09 May 2016 https://doi.org/10.1002/slct.201600369Citations: 6 Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Graphical Abstract Redox partner interaction sites (RPISs) in cytochrome P450 monooxygenases: Most of the cytochrome P450 monooxygenases require a redox partner for catalytic activity. We have analysed available literature, sequences and structures of cytochrome P450s, and identified six RPISs. The findings were applied to design a CYP153 A variant with improved coupling efficiency. Abstract The native redox partners of many novel cytochrome P450 monooxygenases (CYPs) are unknown. Therefore, they are combined with non-native redox partners to obtain catalytically active systems. Understanding the CYP-redox partner interactions is the basis of successful protein engineering. Six redox partner interaction sites (RPISs) were identified by systematic literature, sequence, and structure analyses. All six RPISs are proposed to contribute to class II CYP-redox partner interaction interface, whereas four and five contribute to the interaction interface in class I prokaryotic and mitochondrial CYPs, respectively. The significance of the identified RPISs was tested by designing seven variants of CYP153 A (class I) as a fusion protein with its non-native redox partner CYP102 A1 reductase (class II) and measuring electron coupling efficiencies with a precision of 1–3 %. The best variant K166Q had an improved electron coupling efficiency of 64 % as compared to 53 % for the wild type. Citing Literature Supporting Information As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Filename Description slct201600369-sup-0001-misc_information.pdf161.4 KB Supplementary Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article. Volume1, Issue6May 1, 2016Pages 1243-1251 RelatedInformation