Expanding the Substrate Scope of a Bacterial Nucleotidyltransferase via Allosteric Mutations.
ACS Infect Dis
; 8(10): 2035-2044, 2022 10 14.
Article
em En
| MEDLINE
| ID: mdl-36106727
ABSTRACT
Bacterial glycoconjugates, such as cell surface polysaccharides and glycoproteins, play important roles in cellular interactions and survival. Enzymes called nucleotidyltransferases use sugar-1-phosphates and nucleoside triphosphates (NTPs) to produce nucleoside diphosphate sugars (NDP-sugars), which serve as building blocks for most glycoconjugates. Research spanning several decades has shown that some bacterial nucleotidyltransferases have broad substrate tolerance and can be exploited to produce a variety of NDP-sugars in vitro. While these enzymes are known to be allosterically regulated by NDP-sugars and their fragments, much work has focused on the effect of active site mutations alone. Here, we show that rational mutations in the allosteric site of the nucleotidyltransferase RmlA lead to expanded substrate tolerance and improvements in catalytic activity that can be explained by subtle changes in quaternary structure and interactions with ligands. These observations will help inform future studies on the directed biosynthesis of diverse bacterial NDP-sugars and downstream glycoconjugates.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Açúcares de Nucleosídeo Difosfato
/
Nucleotidiltransferases
Idioma:
En
Revista:
ACS Infect Dis
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Estados Unidos