Abstract
A series of inhibitors have been designed based on 5′-O-sulfamoyl adenosine (AMS) that display tight binding characteristics towards the inhibition of adenylation (A) domains in nonribosomal peptide synthetases (NRPSs). We recently developed an affinity probe for A domains that could be used to facilitate the specific isolation and identification of NRPS modules. Our synthetic probe, which is a biotinylated variant of l-Phe-AMS (l-Phe-AMS-biotin), selectively targets the A domains in NRPS modules that recognize and convert l-Phe to an aminoacyl adenylate in whole proteomes. In this chapter, we describe the design and synthesis of l-Phe-AMS-biotin and provide a summary of our work towards the development of a series of protocols for the specific enrichment of NRPS modules using this probe.
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Acknowledgement
This work was supported in part by a Grant-in Aid for Young Scientists (B) 26750370 (F.I.) and research grants from the Japan Society of the Promotion of Science and the Ministry of Education, Culture, Sports, Science and Technology, Japan (H.K.).
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Ishikawa, F., Kakeya, H. (2016). Affinity Purification Method for the Identification of Nonribosomal Peptide Biosynthetic Enzymes Using a Synthetic Probe for Adenylation Domains. In: Evans, B. (eds) Nonribosomal Peptide and Polyketide Biosynthesis. Methods in Molecular Biology, vol 1401. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3375-4_4
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DOI: https://doi.org/10.1007/978-1-4939-3375-4_4
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