Skip to Content

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

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

Carbohydrate research (2015-05-15)

Ning Wang, Akira Seko, Yoichi Takeda, Yukishige Ito

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.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

Methanol, NMR reference standard

Sigma-Aldrich

Sodium chloride, BioUltra, for molecular biology, ≥99.5% (AT)

Sigma-Aldrich

Sodium chloride solution, BioUltra, for molecular biology, ~5 M in H₂O

Sigma-Aldrich

Sodium chloride, 99.999% trace metals basis

Sigma-Aldrich

Sodium chloride-³⁵Cl, 99 atom % ³⁵Cl

Sigma-Aldrich

Sodium chloride, random crystals, optical grade, 99.9% trace metals basis

Sigma-Aldrich

Sodium chloride solution, 0.85%

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Sodium chloride, AnhydroBeads^™, −10 mesh, 99.999% trace metals basis

Sigma-Aldrich

Sodium chloride solution, 5 M

Sigma-Aldrich

Sodium chloride solution, 5 M in H₂O, BioReagent, for molecular biology, suitable for cell culture

Sigma-Aldrich

Sodium chloride, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)

Sigma-Aldrich

Sodium chloride, tablet

Sigma-Aldrich

Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture

Sigma-Aldrich

Sodium chloride, BioXtra, ≥99.5% (AT)

Sigma-Aldrich

Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%

Sigma-Aldrich

Sodium chloride, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%

Sigma-Aldrich

Sodium chloride, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture

Sigma-Aldrich

Fmoc N-hydroxysuccinimide ester, ≥98.0% (HPLC)

SAFC

Sodium chloride solution, 5 M

Sigma-Aldrich

Methanol-¹²C, 99.95 atom % ¹²C

Sigma-Aldrich

Methanol solution, NMR reference standard, 4% in methanol-d₄ (99.8 atom % D), NMR tube size 3 mm × 8 in.

Supelco

2,5-Dihydroxybenzoic acid, matrix substance for MALDI-MS, >99.0% (HPLC)

Sigma-Aldrich

2,5-Dihydroxybenzoic acid, 98%