Production of complex human glycoproteins in yeast
Production of complex human glycoproteins in yeast
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DOI：
10.1126/science.1088166
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发表时间：
2003-08-29
期刊：
影响因子：
56.9
通讯作者：
Gerngross, TU
中科院分区：
文献类型：
--
作者：
Hamilton, SR;Bobrowicz, P;Gerngross, TU
我们报告了酵母pichia Pastoris中糖基化途径的人性化，以分泌具有均匀复杂N-糖基化的人类糖蛋白。该过程涉及消除内源性酵母糖基化途径，同时适当地定位了五种活性真核蛋白，包括甘露糖苷酶I和II，N-乙酰葡萄糖转移酶I和II，以及尿苷5'-二磷酸盐（UDP）-N-乙酰酸糖苷氨基氨基糖苷。酶的有针对性定位使生成合成的体内糖基化途径，该途径产生了复杂的人类N-聚糖N-乙酰葡萄糖（2） - 甘露糖（3）-n-乙酰葡萄糖胺（2）（2）（3）（Glcnac（3）（3）（3）（3）（3）（3）（3） ）GlcNAC（2））。在真菌宿主中用均质N-聚糖结构产生人糖蛋白的能力是产生治疗性糖蛋白的一步，并且可能成为阐明糖蛋白结构功能关系的工具。
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.