Ternary complex formation with 5-fluoro-2'-deoxyuridylate and the kinetic properties of thymidylate synthase from Lactobacillus leichmannii.
Indian J Biochem Biophys
;
1993 Apr; 30(2): 103-10
Artigo
em Inglês
| IMSEAR
| ID: sea-26278
ABSTRACT
Lactobacillus leichmannii thymidylate synthase (5,10-CH2-H4PteGludUMP C-methyltransferase, EC 2.1.1.45) forms a tight and stable covalently bonded ternary complex with the inhibitor 5-FdUMP in the presence of the cofactor 5,10-CH2-H4-PteGlu. 'Filter assay' employing the radioactive nucleotide ligand showed that 2 moles of FdUMP are bound per mole enzyme during the ternary complex formation with the L. leichmannii dTMP synthase. This is in line with our earlier observation on the Streptococcus faecium thymidylate synthase [Narasimha Rao, K & Kisliuk R L (1983), Proc Natl Acad Sci USA, 80, 916-920]. The enzyme has Km values of 6.3 x 10(-6) M, 8.2 x 10(-5) M and 1.0 x 10(-4) M for dUMP, (dl)-L-H4PteGlu and Mg2+ respectively; Vmax for dUMP, (dl)-L-H4PteGlu and Mg2+ are; 0.55, 0.5 and 1.1 respectively. It has K(i) values of 6.7 x 10(-6) M, 2.2 x 10(-6) M, 5.0 x 10(-5) M and 2.0 x 10(-4) M for FdUMP, dTMP, MTX and Ca2+ respectively. The type of enzyme inhibition with FdUMP, dTMP, MTX and Ca2+ was competitive. dTMP Studies clearly show the 'end product' inhibition of the enzyme.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Conformação Proteica
/
Timidilato Sintase
/
Sítios de Ligação
/
Cinética
/
Fluordesoxiuridilato
/
Lactobacillus
Idioma:
Inglês
Revista:
Indian J Biochem Biophys
Ano de publicação:
1993
Tipo de documento:
Artigo
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