Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis
CY-007 and CY-049 pteridine glycosyltransferases (PGTs) thatdiffer in sugar donor specificity to catalyze either glucose orxylose transfer to tetrahydrobiopterin were studied here touncover the structural determinants necessary for the specificity.The importance of the C-terminal domain and its resi...
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Korean Society for Biochemistry and Molecular Biology
2013-01-01
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doaj-991e5ad09e13471b88c8fd18d8acf50a2020-11-25T00:08:20ZengKorean Society for Biochemistry and Molecular BiologyBMB Reports1976-66961976-670X2013-01-014613740http://dx.doi.org/10.5483/BMBRep.2013.46.1.147Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesisHye-Lim KimAe Hyun KimMi Bi ParkSoo-Woong LeeYoung Shik ParkCY-007 and CY-049 pteridine glycosyltransferases (PGTs) thatdiffer in sugar donor specificity to catalyze either glucose orxylose transfer to tetrahydrobiopterin were studied here touncover the structural determinants necessary for the specificity.The importance of the C-terminal domain and its residues 218and 258 that are different between the two PGTs was assessed viastructure-guided domain swapping or single and dual amino acidsubstitutions. Catalytic activity and selectivity were altered in allthe mutants (2 chimeric and 6 substitution) to accept bothUDP-glucose and UDP-xylose. In addition, the wild typeactivities were improved 1.6-4.2 fold in 4 substitution mutantsand activity was observed towards another substrate UDP-Nacetylglucosaminein all the substitution mutants from CY-007PGT. The results strongly support essential role of the C-terminaldomain and the two residues for catalysis as well as sugar donorspecificity, bringing insight into the structural features of thePGTs. [BMB Reports 2013; 46(1): 37-40]http://bmbreports.org/jbmb/pdf.php?data=MTMwOTI2MTZAcGRmX3JhaW50cmFjZV9sZWV5c0AlNUI0Ni0xJTVEMTMwMTI5MjExNF8lMjgwMzctMDQwJTI5Qk1CXzEyLTE0Ny5wZGY=Domain swappingPteridine glycosyltransferaseSite-directed mutagenesisSubstrate specificityTetrahydrobiopterin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hye-Lim Kim Ae Hyun Kim Mi Bi Park Soo-Woong Lee Young Shik Park |
spellingShingle |
Hye-Lim Kim Ae Hyun Kim Mi Bi Park Soo-Woong Lee Young Shik Park Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis BMB Reports Domain swapping Pteridine glycosyltransferase Site-directed mutagenesis Substrate specificity Tetrahydrobiopterin |
author_facet |
Hye-Lim Kim Ae Hyun Kim Mi Bi Park Soo-Woong Lee Young Shik Park |
author_sort |
Hye-Lim Kim |
title |
Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis |
title_short |
Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis |
title_full |
Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis |
title_fullStr |
Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis |
title_full_unstemmed |
Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis |
title_sort |
altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis |
publisher |
Korean Society for Biochemistry and Molecular Biology |
series |
BMB Reports |
issn |
1976-6696 1976-670X |
publishDate |
2013-01-01 |
description |
CY-007 and CY-049 pteridine glycosyltransferases (PGTs) thatdiffer in sugar donor specificity to catalyze either glucose orxylose transfer to tetrahydrobiopterin were studied here touncover the structural determinants necessary for the specificity.The importance of the C-terminal domain and its residues 218and 258 that are different between the two PGTs was assessed viastructure-guided domain swapping or single and dual amino acidsubstitutions. Catalytic activity and selectivity were altered in allthe mutants (2 chimeric and 6 substitution) to accept bothUDP-glucose and UDP-xylose. In addition, the wild typeactivities were improved 1.6-4.2 fold in 4 substitution mutantsand activity was observed towards another substrate UDP-Nacetylglucosaminein all the substitution mutants from CY-007PGT. The results strongly support essential role of the C-terminaldomain and the two residues for catalysis as well as sugar donorspecificity, bringing insight into the structural features of thePGTs. [BMB Reports 2013; 46(1): 37-40] |
topic |
Domain swapping Pteridine glycosyltransferase Site-directed mutagenesis Substrate specificity Tetrahydrobiopterin |
url |
http://bmbreports.org/jbmb/pdf.php?data=MTMwOTI2MTZAcGRmX3JhaW50cmFjZV9sZWV5c0AlNUI0Ni0xJTVEMTMwMTI5MjExNF8lMjgwMzctMDQwJTI5Qk1CXzEyLTE0Ny5wZGY= |
work_keys_str_mv |
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