Structural and Functional Analyses of the Tridomain‐Nonribosomal Peptide Synthetase FmoA3 for 4‐Methyloxazoline Ring Formation
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- Yohei Katsuyama
- Department of Biotechnology Graduate School of Agricultural and Life science The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku Tokyo 113-8657 Japan
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- Kaoru Sone
- Department of Biotechnology Graduate School of Agricultural and Life science The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku Tokyo 113-8657 Japan
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- Ayaka Harada
- Structural Biology Research Center Institute of Materials Structure Science High Energy Accelerator Research Organization (KEK) 1-1 Oho, Tsukuba Ibaraki 305-0801 Japan
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- Seiji Kawai
- Department of Biotechnology Graduate School of Agricultural and Life science The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku Tokyo 113-8657 Japan
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- Naoki Urano
- Department of Biotechnology Graduate School of Agricultural and Life science The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku Tokyo 113-8657 Japan
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- Naruhiko Adachi
- Structural Biology Research Center Institute of Materials Structure Science High Energy Accelerator Research Organization (KEK) 1-1 Oho, Tsukuba Ibaraki 305-0801 Japan
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- Toshio Moriya
- Structural Biology Research Center Institute of Materials Structure Science High Energy Accelerator Research Organization (KEK) 1-1 Oho, Tsukuba Ibaraki 305-0801 Japan
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- Masato Kawasaki
- Structural Biology Research Center Institute of Materials Structure Science High Energy Accelerator Research Organization (KEK) 1-1 Oho, Tsukuba Ibaraki 305-0801 Japan
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- Kazuo Shin‐ya
- National Institute of Advanced Industrial Science and Technology (AIST) 2-4-7 Aomi, Koto-ku Tokyo 135-0064 Japan
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- Toshiya Senda
- Structural Biology Research Center Institute of Materials Structure Science High Energy Accelerator Research Organization (KEK) 1-1 Oho, Tsukuba Ibaraki 305-0801 Japan
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- Yasuo Ohnishi
- Department of Biotechnology Graduate School of Agricultural and Life science The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku Tokyo 113-8657 Japan
説明
<jats:title>Abstract</jats:title><jats:p>Nonribosomal peptide synthetases (NRPSs) are attractive targets for bioengineering to generate useful peptides. FmoA3 is a single modular NRPS composed of heterocyclization (Cy), adenylation (A), and peptidyl carrier protein (PCP) domains. It uses α‐methyl‐<jats:sc>l</jats:sc>‐serine to synthesize a 4‐methyloxazoline ring, probably with another Cy domain in the preceding module FmoA2. Here, we determined the head‐to‐tail homodimeric structures of FmoA3 by X‐ray crystallography (<jats:italic>apo</jats:italic>‐form, with adenylyl‐imidodiphosphate and α‐methyl‐<jats:sc>l</jats:sc>‐seryl‐AMP) and cryogenic electron microscopy single particle analysis, and performed site‐directed mutagenesis experiments. The data revealed that α‐methyl‐<jats:sc>l</jats:sc>‐serine can be accommodated in the active site because of the extra space around Ala688. The Cy domains of FmoA2 and FmoA3 catalyze peptide bond formation and heterocyclization, respectively. FmoA3’s Cy domain seems to lose its donor PCP binding activity. The collective data support a proposed catalytic cycle of FmoA3.</jats:p>
収録刊行物
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- Angewandte Chemie International Edition
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Angewandte Chemie International Edition 60 (26), 14554-14562, 2021-05-17
Wiley
