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Preparation of asparagine-linked monoglucosylated high-mannose-type oligosaccharide from egg yolk.

Posted by on in 2015
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Wang N1Seko A2Takeda Y2Ito Y3. 2015. Carbohydr Res. 411:37-41. doi: 10.1016/j.carres.2015.04.011. Epub 2015 Apr 26.

1Japan Science and Technology Agency (JST), ERATO, Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Synthetic Cellular Chemistry Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
2Japan Science and Technology Agency (JST), ERATO, Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
3Japan Science and Technology Agency (JST), ERATO, Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Synthetic Cellular Chemistry Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. Electronic address: yukito@riken.jp.

Abstract

Monoglucosylated high-mannose-type glycan (Glc1Man9GlcNAc2: G1M9) is well-known as a key glycoform in the glycoprotein folding process, which is specifically recognized by lectin chaperones calnexin (CNX) and calreticulin (CRT) in the endoplasmic reticulum (ER). In this work, we developed an efficient method for the preparation of G1M9-Asn. The G1M9-Asn was obtained from the IgY-rich fraction derived from hen egg yolk by the digestion with pronase. The α-amino group of asparagine in G1M9-Asn was protected with the 9-fluorenylmethyloxycarbonyl (Fmoc) group and the labeled glycans were subsequently purified using high performance liquid chromatography (HPLC). This method will provide useful substrates for analysis of the glycoprotein folding cycle in the ER.

Copyright © 2015 Elsevier Ltd. All rights reserved.

KEYWORDS:

Egg yolk; Glycopeptide; IgY; Monoglucosylated high-mannose-type oligosaccharide

PMID:
 
25970848
 
[PubMed - in process]
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