Details

Autor(en) / Beteiligte

• 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.

Titel

Production of Complex Human Glycoproteins in Yeast

Ist Teil von

• Science (American Association for the Advancement of Science), 2003-08, Vol.301 (5637), p.1244-1246

Ort / Verlag

Washington, DC: American Association for the Advancement of Science

Erscheinungsjahr

2003

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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$N-acetylglucosamine_{2}-mannose_{3}-N-acetylglucosamine_{2} (GlcNAc_{2}Man_{3}GlcNAc_{2})$. 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.