Document Detail

Cloning and characterization of a viral α2-3-sialyltransferase (vST3Gal-I) for the synthesis of sialyl Lewisx.
MedLine Citation:
PMID:  20978012     Owner:  NLM     Status:  MEDLINE    
Sialyl Lewis(x) (SLe(x), Siaα2-3Galβ1-4(Fucα1-3)GlcNAcβOR) is an important sialic acid-containing carbohydrate epitope involved in many biological processes such as inflammation and cancer metastasis. In the biosynthetic process of SLe(x), α2-3-sialyltransferase-catalyzed sialylation generally proceeds prior to α1-3-fucosyltransferase-catalyzed fucosylation. For the chemoenzymatic synthesis of SLe(x) containing different sialic acid forms, however, it would be more efficient if diverse sialic acid forms are transferred in the last step to the fucosylated substrate Lewis(x) (Le(x)). An α2-3-sialyltransferase obtained from myxoma virus-infected European rabbit kidney RK13 cells (viral α2-3-sialyltransferase (vST3Gal-I)) was reported to be able to tolerate fucosylated substrate Le(x). Nevertheless, the substrate specificity of the enzyme was only determined using partially purified protein from extracts of cells infected with myxoma virus. Herein we demonstrate that a previously reported multifunctional bacterial enzyme Pasteurella multocida sialyltransferase 1 (PmST1) can also use Le(x) as an acceptor substrate, although at a much lower efficiency compared to nonfucosylated acceptor. In addition, N-terminal 30-amino-acid truncated vST3Gal-I has been successfully cloned and expressed in Escherichia coli Origami™ B(DE3) cells as a fusion protein with an N-terminal maltose binding protein (MBP) and a C-terminal His(6)-tag (MBP-Δ30vST3Gal-I-His(6)). The viral protein has been purified to homogeneity and characterized biochemically. The enzyme is active in a broad pH range varying from 5.0 to 9.0. It does not require a divalent metal for its α2-3-sialyltransferase activity. It has been used in one-pot multienzyme sialylation of Le(x) for the synthesis of SLe(x) containing different sialic acid forms with good yields.
Go Sugiarto; Kam Lau; Hai Yu; Stephanie Vuong; Vireak Thon; Yanhong Li; Shengshu Huang; Xi Chen
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2010-10-26
Journal Detail:
Title:  Glycobiology     Volume:  21     ISSN:  1460-2423     ISO Abbreviation:  Glycobiology     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-02-07     Completed Date:  2011-06-28     Revised Date:  2013-07-03    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  387-96     Citation Subset:  IM    
Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Amino Acid Sequence
Base Sequence
Enzyme Assays
Hydrogen-Ion Concentration
Molecular Sequence Data
Myxoma virus / enzymology*
Oligosaccharides / biosynthesis*,  chemistry
Recombinant Fusion Proteins / biosynthesis*,  chemistry,  isolation & purification
Sequence Alignment
Sialyltransferases / biosynthesis*,  chemistry,  isolation & purification
Grant Support
Reg. No./Substance:
0/5-acetylneuraminyl-(2-3)-galactosyl-(1-4)-(fucopyranosyl-(1-3))-N-acetylglucosamine; 0/Oligosaccharides; 0/Recombinant Fusion Proteins; EC 2.4.99.-/Sialyltransferases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Overexpression of Man2C1 leads to protein underglycosylation and upregulation of endoplasmic reticul...
Next Document:  Changing trends and predictors of outcome in patients with acute poisoning admitted to the intensive...