Laboratory for Bioorganic Chemistry, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
Cell Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
Science. 2013 Dec 13;342(6164):1357-1360. doi: 10.1126/science.1245095.
Erythropoietin is a signaling glycoprotein that controls the fundamental process of erythropoiesis, orchestrating the production and maintenance of red blood cells. As administrated clinically, erythropoietin has a polypeptide backbone with complex dishomogeneity in its carbohydrate domains. Here we describe the total synthesis of homogeneous erythropoietin with consensus carbohydrate domains incorporated at all of the native glycosylation sites. The oligosaccharide sectors were built by total synthesis and attached stereospecifically to peptidyl fragments of the wild-type primary sequence, themselves obtained by solid-phase peptide synthesis. The glycopeptidyl constructs were joined by chemical ligation, followed by metal-free dethiylation, and subsequently folded. This homogeneous erythropoietin glycosylated at the three wild-type aspartates with N-linked high-mannose sialic acid-containing oligosaccharides and O-linked glycophorin exhibits Procrit-level in vivo activity in mice.
促红细胞生成素是一种信号糖蛋白，它控制着红细胞生成的基本过程，协调红细胞的生成和维持。在临床上使用时，促红细胞生成素具有一个多肽主链，其碳水化合物结构域具有复杂的不均一性。在这里，我们描述了具有共识碳水化合物结构域的均一促红细胞生成素的全合成，这些结构域在所有天然糖基化位点都被掺入。寡糖部分通过全合成构建，并特异性地连接到野生型一级序列的肽片段上，这些肽片段本身是通过固相肽合成获得的。糖肽构建体通过化学连接，然后进行无金属脱硫，随后进行折叠。这种在三个野生型天冬氨酸上进行 N 连接的高甘露糖唾液酸化含有寡糖和 O 连接的糖蛋白的均一促红细胞生成素在小鼠体内具有 Procrit 水平的活性。
