Methods for Studying the Radical SAM Enzymes in Diphthamide Biosynthesis.
Methods Enzymol
; 606: 421-438, 2018.
Article
em En
| MEDLINE
| ID: mdl-30097101
ABSTRACT
Diphthamide is a unique posttranslational modification on translation elongation factor 2 (EF2) in archaea and eukaryotes. Biosynthesis of diphthamide was proposed to involve four steps. The first step is a CC bond forming reaction catalyzed by unique radical S-adenosylmethionine (SAM) enzymes. Classical radical SAM enzymes use SAM and [4Fe-4S] clusters to generate a 5'-deoxyadenynal radical and catalyze numerous reactions. Radical SAM enzymes in diphthamide biosynthesis cleave a different CS bond in SAM to generate a 3-amino-3-carboxypropyl radical and modify a histidine residue of substrate protein EF2. Here, we describe our investigations on these unique radical SAM enzymes, including the preparation, characterization, and activity assays we have developed.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
S-Adenosilmetionina
/
Alquil e Aril Transferases
/
Proteínas Arqueais
/
Ensaios Enzimáticos
/
Histidina
Idioma:
En
Revista:
Methods Enzymol
Ano de publicação:
2018
Tipo de documento:
Article