Publication Date:
2003-08-30
Description:
We report the humanization of the glycosylation pathway in the yeast Pichia pastoris to secrete a human glycoprotein with uniform complex N-glycosylation. The process involved eliminating endogenous yeast glycosylation pathways, while properly localizing five active eukaryotic proteins, including mannosidases I and II, N-acetylglucosaminyl transferases I and II, and uridine 5'-diphosphate (UDP)-N-acetylglucosamine transporter. Targeted localization of the enzymes enabled the generation of a synthetic in vivo glycosylation pathway, which produced the complex human N-glycan N-acetylglucosamine2-mannose3-N-acetylglucosamine2 (GlcNAc2Man3GlcNAc2). The ability to generate human glycoproteins with homogeneous N-glycan structures in a fungal host is a step toward producing therapeutic glycoproteins and could become a tool for elucidating the structure-function relation of glycoproteins.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Hamilton, Stephen R -- Bobrowicz, Piotr -- Bobrowicz, Beata -- Davidson, Robert C -- Li, Huijuan -- Mitchell, Teresa -- Nett, Juergen H -- Rausch, Sebastian -- Stadheim, Terrance A -- Wischnewski, Harry -- Wildt, Stefan -- Gerngross, Tillman U -- 1R43GM66690-1/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 2003 Aug 29;301(5637):1244-6.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Thayer School of Engineering and the Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/12947202" target="_blank"〉PubMed〈/a〉
Keywords:
Animals
;
Catalytic Domain
;
Endoplasmic Reticulum/metabolism
;
*Genetic Engineering
;
Glycoproteins/*biosynthesis/chemistry/genetics
;
Glycosylation
;
Golgi Apparatus/metabolism
;
Humans
;
Mannosidases/*genetics/metabolism
;
Membrane Transport Proteins/metabolism
;
N-Acetylglucosaminyltransferases/metabolism
;
Peptide Library
;
Pichia/enzymology/*genetics/metabolism
;
Polysaccharides/chemistry/*metabolism
;
Protein Processing, Post-Translational
;
Protein Transport
;
Recombinant Fusion Proteins/metabolism
;
Recombinant Proteins/*biosynthesis
;
Transformation, Genetic
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
Permalink