Altered sugar donor specificity and catalytic activity of pteridine glycosyltransferases by domain swapping or site-directed mutagenesis

被引:7
|
作者
Kim, Hye-Lim [1 ]
Kim, Ae Hyun [1 ]
Park, Mi Bi [1 ]
Lee, Soo-Woong [2 ]
Park, Young Shik [1 ]
机构
[1] Inje Univ, Sch Biol Sci, Kimhae 621749, South Korea
[2] Inje Univ, Adv Res Ctr Multiple Myeloma, Coll Med, Pusan 614735, South Korea
关键词
Domain swapping; Pteridine glycosyltransferase; Site-directed mutagenesis; Substrate specificity; Tetrahydrobiopterin; FAMILY GT4; TETRAHYDROBIOPTERIN; GLUCOSYLTRANSFERASE; GLUCOSE;
D O I
10.5483/BMBRep.2013.46.1.147
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 domain and 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 activity and 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 donor specificity, bringing insight into the structural features of the PGTs. [BMB Reports 2013; 46(1): 37-40]
引用
收藏
页码:37 / 40
页数:4
相关论文
共 50 条
  • [21] ALTERATION OF THE ENZYMATIC SPECIFICITY OF HUMAN ANGIOGENIN BY SITE-DIRECTED MUTAGENESIS
    CURRAN, TP
    SHAPIRO, R
    RIORDAN, JF
    BIOCHEMISTRY, 1993, 32 (09) : 2307 - 2313
  • [22] Effect on product specificity of cyclodextrin glycosyltransferase by site-directed mutagenesis
    Kim, YH
    Bae, KH
    Kim, TJ
    Park, KH
    Lee, HS
    Byun, SM
    BIOCHEMISTRY AND MOLECULAR BIOLOGY INTERNATIONAL, 1997, 41 (02): : 227 - 234
  • [23] Alteration of substrate specificity of leucine dehydrogenase by site-directed mutagenesis
    Kataoka, K
    Tanizawa, K
    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 2003, 23 (2-6) : 299 - 309
  • [24] AN INCREASE IN THE TRANSGLYCOSYLATION ACTIVITY OF SACCHAROMYCOPSIS ALPHA-AMYLASE ALTERED BY SITE-DIRECTED MUTAGENESIS
    MATSUI, I
    ISHIKAWA, K
    MIYAIRI, S
    FUKUI, S
    HONDA, K
    BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1077 (03) : 416 - 419
  • [25] SITE-DIRECTED MUTAGENESIS TO FINE-TUNE ENZYME SPECIFICITY
    UEMURA, H
    ROGERS, MJ
    SWANSON, R
    WATSON, L
    SOLL, D
    PROTEIN ENGINEERING, 1988, 2 (04): : 293 - 296
  • [26] Analysis of human ITPase nucleobase specificity by site-directed mutagenesis
    Gall, Amanda D.
    Gall, Anthony
    Moore, Ashley C.
    Aune, Martin K.
    Heid, Steven
    Mori, Ayaka
    Burgis, Nicholas E.
    BIOCHIMIE, 2013, 95 (09) : 1711 - 1721
  • [27] USE OF SITE-DIRECTED MUTAGENESIS TO INVESTIGATE THE BASIS FOR THE SPECIFICITY OF HIRUDIN
    BRAUN, PJ
    DENNIS, S
    HOFSTEENGE, J
    STONE, SR
    BIOCHEMISTRY, 1988, 27 (17) : 6517 - 6522
  • [28] DETERMINATION OF THE CATALYTIC SITE OF CREATINE-KINASE BY SITE-DIRECTED MUTAGENESIS
    LIN, LJ
    PERRYMAN, MB
    FRIEDMAN, D
    ROBERTS, R
    MA, TS
    BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1994, 1206 (01): : 97 - 104
  • [29] SITE-DIRECTED MUTAGENESIS OF THE CATALYTIC SITE OF F1-ATPASE
    PARSONAGE, D
    SENIOR, AE
    BIOPHYSICAL JOURNAL, 1987, 51 (02) : A242 - A242
  • [30] Site-directed mutagenesis of yeast eEF1A; viable mutants with altered nucleotide specificity.
    Cavallius, J
    Merrick, WC
    FASEB JOURNAL, 1997, 11 (09): : A1403 - A1403