Direct assay for endo-α-mannosidase substrate preference on correctly folded and misfolded model glycoproteins
| Autor: | Masayuki Izumi, Yukishige Ito, Akiko Kanamori, Simone Dedola, Yutaka Makimura, Yoichi Takeda, Akira Seko, Yasuhiro Kajihara |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Mannosidase Protein Folding Mutant UGGT macromolecular substances Biochemistry Substrate Specificity Analytical Chemistry 03 medical and health sciences symbols.namesake Protein structure alpha-Mannosidase Glycoproteins Plant Proteins chemistry.chemical_classification Molecular Structure 030102 biochemistry & molecular biology biology Organic Chemistry General Medicine Golgi apparatus carbohydrates (lipids) Folding (chemistry) 030104 developmental biology chemistry Mutation biology.protein symbols lipids (amino acids peptides and proteins) Glucosyltransferase Glycoprotein |
| Zdroj: | Carbohydrate Research. 434:94-98 |
| ISSN: | 0008-6215 |
| DOI: | 10.1016/j.carres.2016.08.008 |
| Popis: | We previously reported a unique assay system for UDP-glucose glycoprotein glucosyltransferase (UGGT) toward glycoprotein folding intermediates during the folding process. The assay involved the in vitro folding of both high-mannose type oligosaccharyl crambin, which yielded only the correctly folded glycoprotein form (M9-glycosyl-native-crambin), and its mutant, which yielded misfolded glycoproteins (M9-glycosyl-misfolded-crambin), in the presence of UGGT. The process successfully yielded both mono-glucosylated M9-glycosyl-native-crambin (G1M9-glycosyl-native-crambin) and M9-glycosyl-misfolded-crambin (G1M9-glycosyl-misfolded-crambin). Here, we report the use of our in vitro folding system to evaluate the substrate preference of Golgi endo-α-mannosidase against G1M9-native and -misfolded glycoprotein forms. In our assay Golgi endo-α-mannosidase removed Glc-α-1-3-Man unit from G1M9-native and -misfolded-crambins clearly proving that Golgi endo-α-mannosidase does not have specific preference for correctly folded or misfolded protein structure. |
| Databáze: | OpenAIRE |
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