Biochemical characterization of the pneumococcal glucose 1-phosphate uridylyltransferase (GalU) essential for capsule biosynthesis.
The glucose 1-phosphate uridylyltransferase (GalU) is absolutely required for the biosynthesis of capsular polysaccharide, the sine qua non virulence factor of Streptococcus pneumoniae. The pneumococcal GalU protein was overexpressed in Escherichia coli, and purified. GalU showed a pI of 4.23, and catalyzed the reversible formation of UDP-glucose and pyrophosphate from UTP and glucose 1-phosphate with K(m) values of 0.4 mM: for UDP-glucose, 0.26 mM: for pyrophosphate, 0.19 mM: for glucose 1-phosphate, and 0.24 mM: for UTP. GalU has an optimum pH of 8-8.5, and requires Mg(2+) for activity. Neither ADP-glucose nor TDP-glucose is utilized as substrates in vitro. The purification of GalU represents a fundamental step to provide insights on drug design to control the biosynthesis of the main pneumococcal virulence factor.
Bonofiglio
Laura
L
Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Argentina.
García
Ernesto
E
Mollerach
Marta
M
D016667:Bacterial Capsules / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000235:genetics; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000378:metabolism*
D011135:Polysaccharides, Bacterial; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase