Details

Autor(en) / Beteiligte

• Iwamoto, Shogo
• Kasahara, Yuta
• Kamei, Ken-ichi
• Seko, Akira
• Takeda, Yoichi
• Ito, Yukishige
• Matsuo, Ichiro

Titel

Measurement of endo-α-mannosidase activity using a fluorescently labeled oligosaccharide derivative

Ist Teil von

• Bioscience, biotechnology, and biochemistry, 2014, Vol.78 (6), p.927-936

Ort / Verlag

England: Taylor & Francis

Erscheinungsjahr

2014

Quelle

MEDLINE

Beschreibungen/Notizen

• Endo-α-mannosidase, a GH99-family glycoside hydrolase, cleaves α-mannoside linkages with glucose residues. This enzyme is proposed to play a critical role in N-glycan processing for deglucosylation. To measure endo-α-mannosidase activity, we synthesized a fluorescently labeled tetrasaccharide derivative (Glcα1-3Manα1-2Manα1-2Manα1-O-C 3 H 6 -NH-Dansyl) in a stereocontrolled manner. The tetrasaccharide skeleton was prepared by step-wise coupling using mannose donors 4 and 7. The 1,2-cis α-glycosidic linkage on the non-reducing end of the glucose residue was constructed by inversion of the stereochemistry of the C-2 hydroxyl group in the α-mannose residue. Finally, the dansyl group was introduced at the reducing end via an aminopropyl linker. This probe successfully measured endo-α-mannosidase activity. A fluorescently labeled tetrasaccharide 1 (Glcα1-3Manα1-2Manα1-2Manα1-O-C 3 H 6 -NH-Dansyl) was chemically synthesized. Using this probe, we successfully measured endo-α-mannosidase activity.