Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
Department of Biochemistry, Universität Bayreuth, Germany.
Angew Chem Int Ed Engl. 2017 May 2;56(19):5252-5257. doi: 10.1002/anie.201701362. Epub 2017 Apr 5.
The main glycoforms of the hydrophobic lysosomal glycoprotein saposin D (SapD) were synthesized by native chemical ligation. An approach for the challenging solid-phase synthesis of the fragments was developed. Three SapD glycoforms were obtained following a general and robust refolding and purification protocol. A crystal structure of one glycoform confirmed its native structure and disulfide pattern. Functional assays revealed that the lipid-binding properties of three SapD glycoforms are highly affected by the single sugar moiety of SapD showing a dependency of the size and the type of N-glycan.
疏水溶酶体糖蛋白 saposin D（SapD）的主要糖型通过天然化学连接合成。开发了一种用于挑战性固相合成片段的方法。采用通用且稳健的复性和纯化方案获得了三种 SapD 糖型。一种糖型的晶体结构证实了其天然结构和二硫键模式。功能测定表明，三种 SapD 糖型的脂质结合特性受 SapD 中单糖部分的强烈影响，表现出对 N-聚糖大小和类型的依赖性。
