The kinetic mechanism of S. pneumoniae DNA ligase and inhibition by adenosine-based antibacterial compounds.
Biochem Pharmacol
; 84(5): 654-60, 2012 Sep 01.
Article
em En
| MEDLINE
| ID: mdl-22743594
ABSTRACT
The NAD-dependent DNA ligase is an excellent target for the discovery of antibacterial agents with a novel mode of action. In this work the DNA ligase from Streptococcus pneumoniae was investigated for its steady-state kinetic parameters and inhibition by compounds with an adenosine substructure. Inhibition by substrate DNA that was observed in the enzyme turnover experiments was verified by direct binding measurements using isothermal titration calorimetry (ITC). The substrate-inhibited enzyme form was identified as deadenylated DNA ligase. The binding potencies of 2-(butylsulfanyl) adenosine and 2-(cyclopentyloxy) adenosine were not significantly affected by the presence of the enzyme-bound DNA substrate. Finally, a mutant protein was prepared that was known to confer resistance to the adenosine compounds' antibacterial activity. The mutant protein was shown to have little catalytic impairment yet it was less susceptible to adenosine compound inhibition.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Ligases
/
Streptococcus pneumoniae
/
Adenosina
/
Inibidores Enzimáticos
/
Antibacterianos
Idioma:
En
Revista:
Biochem Pharmacol
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos