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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

要旨

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.

材料

製品番号

ブランド

製品内容

Sigma-Aldrich

塩化ナトリウム, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)

Sigma-Aldrich

塩化ナトリウム溶液, 0.9% in water, BioXtra, suitable for cell culture

Sigma-Aldrich

塩化ナトリウム, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%

Sigma-Aldrich

塩化ナトリウム溶液, 5 M in H₂O, BioReagent, for molecular biology, suitable for cell culture

Sigma-Aldrich

塩化ナトリウム, JIS special grade, ≥99.5%

Sigma-Aldrich

メタノール, JIS special grade, ≥99.8%

Sigma-Aldrich

塩化ナトリウム溶液, BioUltra, for molecular biology, ~5 M in H₂O

Sigma-Aldrich

塩化ナトリウム, 99.999% trace metals basis

Sigma-Aldrich

2,5-ジヒドロキシ安息香酸, 98%

Sigma-Aldrich

塩化ナトリウム, BioUltra, for molecular biology, ≥99.5% (AT)

Supelco

2,5-ジヒドロキシ安息香酸, matrix substance for MALDI-MS, >99.0% (HPLC)

Sigma-Aldrich

メタノール, SAJ first grade, ≥99.5%

Sigma-Aldrich

Fmoc N-ヒドロキシスクシンイミドエステル, ≥98.0% (HPLC)

Sigma-Aldrich

メタノール, anhydrous, 99.8%

Sigma-Aldrich

塩化ナトリウム, SAJ first grade, ≥99.0%

Sigma-Aldrich

塩化ナトリウム, BioXtra, ≥99.5% (AT)

Sigma-Aldrich

塩化ナトリウム, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture

Sigma-Aldrich

メタノール, suitable for HPLC

Sigma-Aldrich

メタノール, SAJ special grade

Sigma-Aldrich

塩化ナトリウム, AnhydroBeads^™, −10 mesh, 99.999% trace metals basis

Sigma-Aldrich

塩化ナトリウム-³⁵Cl, 99 atom % ³⁵Cl

Sigma-Aldrich

塩化ナトリウム, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%

Sigma-Aldrich

塩化ナトリウム溶液, 1 M

Sigma-Aldrich

塩化ナトリウム溶液, 5 M

Sigma-Aldrich

塩化ナトリウム, random crystals, optical grade, 99.9% trace metals basis

Sigma-Aldrich

メタノール, NMR reference standard

Sigma-Aldrich

塩化ナトリウム溶液, 0.85%

Sigma-Aldrich

塩化ナトリウム溶液, 0.1 M

Sigma-Aldrich

Methanol-¹²C, 99.95 atom % ¹²C

Supelco

メタノール溶液, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%