|
KMID : 0624620130460010037
|
|
BMB Reports
2013 Volume.46 No. 1 p.37 ~ p.40
|
|
|
Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis
|
|
Kim Hye-Lim
Kim Ae-Hyun
Park Mi-Bi
Lee Soo-Woong
Park Young-Shik
|
|
|
Abstract
|
|
|
|
CY-007 and CY-049 pteridine glycosyltransferases (PGTs) that differ in sugar donor specificity to catalyze either glucose or xylose transfer to tetrahydrobiopterin were studied here to uncover the structural determinants necessary for the specificity. The importance of the C-terminal domainand its residues 218 and 258 that are different between the two PGTs was assessed via structure-guided domain swapping or single and dual amino acid substitutions. Catalytic activityand selectivity were altered in all the mutants (2 chimeric and 6 substitution) to accept both UDP-glucose and UDP-xylose. In addition, the wild type activities were improved 1.6-4.2 fold in 4 substitution mutants and activity was observed towards another substrate UDP-N-acetylglucosamine in all the substitution mutants from CY-007 PGT. The results strongly support essential role of the C-terminal domain and the two residues for catalysis as well as sugar donorspecificity, bringing insight into the structural features of the PGTs.
|
|
|
KEYWORD
|
|
|
Domain swapping, Pteridine glycosyltransferase, Site-directed mutagenesis, Substrate specificity, Tetrahydrobiopterin
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|