Journal Article10.1021/ACSCHEMBIO.0C00637
Non-Heme Monooxygenase ThoJ Catalyzes Thioholgamide β-Hydroxylation.
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TL;DR: The construction of a heterologous host capable of producing thioholgamide with a 15-fold increased yield compared to the wild-type strain is reported, enabling the dissection of the individual biosynthetic steps involved in biosynthesis of this exciting RiPP family.
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Abstract: Thioviridamide-like compounds, including thioholgamides, are ribosomally synthesized and post-translationally modified peptide natural products with potent anticancer cell activity and an unprecedented structure. Very little is known about their biosynthesis, and we were intrigued by the β-hydroxy-N1, N3-dimethylhistidinium moiety found in these compounds. Here we report the construction of a heterologous host capable of producing thioholgamide with a 15-fold increased yield compared to the wild-type strain. A knockout of thoJ, encoding a predicted nonheme monooxygenase, shows that ThoJ is essential for thioholgamide β-hydroxylation. The crystal structure of ThoJ exhibits a typical mono/dioxygenase fold with conserved key active-site residues. Yet, ThoJ possesses a very large substrate binding pocket that appears suitable to receive a cyclic thioholgamide intermediate for hydroxylation. The improved production of the heterologous host will enable the dissection of the individual biosynthetic steps involved in biosynthesis of this exciting RiPP family.
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Citations
Total In Vitro Biosynthesis of the Thioamitide Thioholgamide and Investigation of the Pathway.
TL;DR: The complete in vitro reconstitution of the biosynthetic pathway is reported, demonstrating that thioamidation is the first step and acts as a gatekeeper for downstream processing.
Understanding Thioamitide Biosynthesis Using Pathway Engineering and Untargeted Metabolomics
TL;DR: Engineering the thiostreptamide S4 gene cluster in a heterologous host could provide insights into its biosynthesis when coupled with untargeted metabolomics and targeted mutations of the precursor peptide, paving the way for future RiPP discovery and engineering.
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Biochemical Reconstitution Reveals the Biosynthetic Timing and Substrate Specificity for Thioamitides.
Jiang Xiong,Shangwen Luo,Cheng-Xiao Qin,Jiaojiao Cui,Yu-xia Ma,Meng-Xue Guo,Shaozhong Zhang,Yan Li,Kun Gao,Shi-Hui Dong +9 more
TL;DR: This study reconstituted thioamitide key PTMs through in vitro enzymatic reactions and gene coexpressions in E. coli and rigorously demonstrated the order of those modifications.
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Tryptophan-Centric Bioinformatics Identifies New Lasso Peptide Modifications.
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References
The Protein Data Bank
Helen M. Berman,John D. Westbrook,Zukang Feng,Gary L. Gilliland,Talapady N. Bhat,Helge Weissig,Ilya N. Shindyalov,Philip E. Bourne +7 more
TL;DR: The goals of the PDB are described, the systems in place for data deposition and access, how to obtain further information and plans for the future development of the resource are described.
ConSurf 2016: an improved methodology to estimate and visualize evolutionary conservation in macromolecules.
TL;DR: Several new features into ConSurf are introduced, including automatic selection of the best evolutionary model used to infer the rates, the able to homology-model query proteins, prediction of the secondary structure of query RNA molecules from sequence, the ability to view the biological assembly of a query (in addition to the single chain), mapping of the conservation grades onto 2D RNA models and an advanced view of the phylogenetic tree.
ConSurf 2010: calculating evolutionary conservation in sequence and structure of proteins and nucleic acids
TL;DR: The new version of the ConSurf web server is presented, providing an easier and more intuitive step-by-step interface, while offering the user more flexibility during the process, and calculates the evolutionary rates for nucleic acid sequences.
Ribosomally synthesized and post-translationally modified peptide natural products: Overview and recommendations for a universal nomenclature
Paul G. Arnison,Mervyn J. Bibb,Gabriele Bierbaum,Albert A. Bowers,Tim S. Bugni,Grzegorz Bulaj,Julio A. Camarero,Dominic J. Campopiano,Gregory L. Challis,Jon Clardy,Paul D. Cotter,David J. Craik,Michael J. Dawson,Elke Dittmann,Stefano Donadio,Pieter C. Dorrestein,K. D. Entian,Michael A. Fischbach,John S. Garavelli,Ulf Göransson,Christian W. Gruber,Daniel H. Haft,Thomas Hemscheidt,Christian Hertweck,Colin Hill,Alexander R. Horswill,Marcel Jaspars,Wendy L. Kelly,Judith P. Klinman,Oscar P. Kuipers,A. James Link,Wen Liu,Mohamed A. Marahiel,Douglas A. Mitchell,Gert N. Moll,Bradley S. Moore,Rolf Müller,Satish K. Nair,Ingolf F. Nes,Gillian E. Norris,Baldomero M. Olivera,Hiroyasu Onaka,Mark L. Patchett,Joern Piel,Joern Piel,Martin J. T. Reaney,Sylvie Rebuffat,R. Paul Ross,Hans-Georg Sahl,Eric W. Schmidt,Michael E. Selsted,Konstantin Severinov,Ben Shen,Kaarina Sivonen,Leif Smith,Torsten Stein,Roderich D. Süssmuth,John R. Tagg,Gong-Li Tang,Andrew W. Truman,John C. Vederas,Christopher T. Walsh,Jonathan D. Walton,Silke C. Wenzel,Joanne M. Willey,Wilfred A. van der Donk +65 more
TL;DR: This review presents recommended nomenclature for the biosynthesis of ribosomally synthesized and post-translationally modified peptides (RiPPs), a rapidly growing class of natural products.
1.8K
ConSurf 2005: the projection of evolutionary conservation scores of residues on protein structures
TL;DR: This new version of ConSurf includes an empirical Bayesian method for scoring conservation, which is more accurate than the maximum-likelihood method that was used in the earlier release and includes a measure of confidence for the inferred amino acid conservation scores.
1.4K