Cross-Linking of the Nonribosomal Peptide Synthetase Adenylation Domain with a Carrier Protein Using a Pantetheine-Type Probe.

Autor:	Miyanaga A; Department of Chemistry, Tokyo Institute of Technology, Tokyo, Japan. miyanaga.a.aa@m.titech.ac.jp., Kudo F; Department of Chemistry, Tokyo Institute of Technology, Tokyo, Japan., Eguchi T; Department of Chemistry, Tokyo Institute of Technology, Tokyo, Japan. eguchi@chem.titech.ac.jp.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2670, pp. 207-217.
DOI:	10.1007/978-1-0716-3214-7_10
Abstrakt:	Adenylation domains (A-domains) are responsible for the selective incorporation of carboxylic acid substrates in the biosynthesis of nonribosomal peptides and related natural products. The A-domain transfers an acyl substrate onto its cognate carrier protein (CP). The proper interactions between an A-domain and the cognate CP are important for functional substrate transfer. To stabilize the transient interactions sufficiently for structural analysis of A-domain-CP complex, vinylsulfonamide adenosine inhibitors have been traditionally used as molecular probes. Recently, we have developed an alternative strategy using a synthetic pantetheine-type probe that enables site-specific cross-linking between an A-domain and a CP. In this chapter, we describe the laboratory protocols for this cross-linking reaction. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu