Document Detail


Metabolic functionalization of recombinant glycoproteins.
MedLine Citation:
PMID:  15379575     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Glycoproteins are essential for cellular communication and are the most rapidly growing class of therapeutic agents. Chemical modification of glycoproteins has been employed to improve their in vivo efficacy or to label them for detection. Methods for the controlled derivatization of glycoproteins are presently limited by the repertoire of natural amino acid side chain and carbohydrate functionalities. Here, we use metabolic oligosaccharide engineering to introduce a bioorthogonal functional group, the azide, into cellular and recombinant glycoproteins for subsequent chemical elaboration via Staudinger ligation. As most therapeutic glycoproteins are sialylated and require this saccharide for optimal pharmacokinetics, we targeted sialic acid as a host for azides using N-azidoacetylmannosamine (ManNAz) as a biosynthetic precursor. Metabolic conversion of ManNAz to N-azidoacetylsialic acid (SiaNAz) within membrane-bound and secreted glycoproteins was quantified in a variety of cell types. SiaNAz was found to comprise between 4% and 41% of total sialosides, depending on the system. Metabolic labeling of recombinant interferon-beta and GlyCAM-Ig was achieved, demonstrating the utility of the method for functionalizing N-linked and O-linked glycoproteins of therapeutic interest. More generally, the generation of recombinant glycoproteins containing chemical handles within their glycans provides a means for studying their behavior and for improving their in vivo efficacy.
Authors:
Sarah J Luchansky; Sulabha Argade; Bradley K Hayes; Carolyn R Bertozzi
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biochemistry     Volume:  43     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  2004 Sep 
Date Detail:
Created Date:  2004-09-21     Completed Date:  2004-11-09     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  12358-66     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of California, Berkeley, California 94720, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Azides / chemistry,  metabolism,  pharmacology
Cell Line
Cercopithecus aethiops
Chromatography, High Pressure Liquid
Cricetinae
Gene Expression
Glycoproteins / chemistry,  genetics,  metabolism*
Humans
Mass Spectrometry
Molecular Structure
N-Acetylneuraminic Acid / metabolism
Oligosaccharides / chemistry,  metabolism
Recombinant Proteins / chemistry,  genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
GM58867/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Azides; 0/Glycoproteins; 0/Oligosaccharides; 0/Recombinant Proteins; 131-48-6/N-Acetylneuraminic Acid

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


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