Use of an unnatural amino acid to map helicase/DNA interfaces via photoactivated crosslinking

Autor:	Alexander T. Duckworth, James L. Keck
Rok vydání:	2022
Předmět:	DNA Replication Escherichia coli Proteins DNA Helicases DNA Amino Acids Article
Zdroj:	Methods in Enzymology ISBN: 9780323914765 Methods Enzymol
DOI:	10.1016/bs.mie.2022.02.019
Popis:	Formation of protein/nucleic acid complexes is essential for life. From DNA replication and repair to transcription and translation, myriad different proteins bind nucleic acids to execute their essential cellular functions. Our understanding of the mechanisms underlying recognition and processing of nucleic acids can be greatly informed by mapping protein domains and residues that form interfaces with their DNA or RNA targets. Here we describe a crosslinking protocol in which the unnatural amino acid p-benzoyl-l-phenylalanine (Bpa) integrated at selected sites within the PriA DNA helicase is used to map surfaces of the protein that interact with specific positions in a synthetic DNA replication fork in vitro.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::71743a5833971d27a82c21a6d258fc16 https://doi.org/10.1016/bs.mie.2022.02.019 Zobrazit plný text záznamu