Glycoproteins in non-native conformations are often toxic to cells and may cause diseases, thus the quality control (QC) system eliminates these unwanted species. Lectin chaperone calreticulin and glucosidase II, both of which recognize the Glc1 Man9 oligosaccharide on glycoproteins, are important components of the glycoprotein QC system. Reported herein is the preparation of Glc1 Man9 -glycoproteins in both native and non-native conformations by using the following sequence: misfolding of chemically synthesized Man9 -glycoprotein, enzymatic glucosylation, and another misfolding step. By using synthetic glycoprotein probes, calreticulin was found to bind preferentially to a hydrophobic non-native glycoprotein whereas glucosidase II activity was not affected by glycoprotein conformation. The results demonstrate the ability of chemical synthesis to deliver homogeneous glycoproteins in several non-native conformations for probing the glycoprotein QC system.

Glycoproteins in non-native conformations are often toxic to cells and may cause diseases,t hus the quality control (QC) system eliminates these unwanted species.Lectin chaperone calreticulin and glucosidase II, both of which recognizet he Glc 1 Man 9 oligosaccharide on glycoproteins,a re important components of the glycoprotein QC system. Reported herein is the preparation of Glc 1 Man 9 -glycoproteins in both native and non-native conformations by using the following sequence:m isfolding of chemically synthesized Man 9 -glycoprotein, enzymatic glucosylation, and another misfolding step.B yu sing synthetic glycoprotein probes, calreticulin was found to bind preferentially to ahydrophobic non-native glycoprotein whereas glucosidase II activity was not affected by glycoprotein conformation. The results demonstrate the ability of chemical synthesis to deliver homogeneous glycoproteins in several non-native conformations for probing the glycoprotein QC system.