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ID 56096
FullText URL
Author
Uemura, Ryota Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science , Okayama University
Ogura, Mikako Faculty of Agriculture , Okayama University
Matsumaru, Chihiro Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science , Okayama University
Akiyama, Tsuyoshi Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science , Okayama University
Maeda, Megumi Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science , Okayama University Kaken ID
Kimura, Yoshinobu Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science , Okayama University Kaken ID
Abstract
Acidic peptide:N-glycanase (aPNGase) plays a pivotal role in plant glycoprotein turnover. For the construction of aPNGase-knockout or -overexpressing plants, a new method to detect the activity in crude plant extracts is required because endogenous peptidases present in the extract hamper enzyme assays using fluorescence-labeled N-glycopeptides as a substrate. In this study, we developed a new method for measuring aPNGase activity in crude extracts from plant materials
Keywords
Acidic PNGase
FNG: free N-glycan
Fuc: L-fucose
Gal: D-galactose
GlcNAc: N-acetyl-D-glucosamine
HPLC: high-performance liquid chromatography
Man: D-mannose
NeuNAc2Gal2GlcNAc2Man3GlcNAc1: NeuNAcα2–6Galβ1–4GlcNAcβ1–2Manα1–6(NeuNAcα2–6Galβ1–4GlcNAcβ1–2Manα1–3)Manβ1–4GlcNAc
NeuNAc2Gal2GlcNAc2Man3GlcNAc2: NeuNAcα2–6Galβ1–4GlcNAcβ1–2Manα1–6(NeuNAcα2–6Galβ1–4GlcNAcβ1–2Manα1–3)Manβ1–4GlcNAcβ1–4GlcNAc
NeuNAc: N-acetylneuraminic acid
PA-: pyridylamino
PNGase-A: aPNGase from almond seed
PNGase-Le: aPNGase from tomato (Solanum lycopersium L.)
PNGase: peptide:N-glycanase
PTC: plant complex type
RCA120: Ricinus communis agglutinin (120 kDa)
RP-HPLC: reversed-phase HPLC
SF-HPLC: size-fractionation HPLC
Xyl: D-xylose
affinity chromatography
enzyme assay
free N-glycans
transgenic plant
Published Date
2018-04
Publication Title
Bioscience, Biotechnology, and Biochemistry
Volume
volume82
Issue
issue7
Publisher
Taylor & Francis
Start Page
1172
End Page
1175
ISSN
09168451
NCID
AA10824164
Content Type
Journal Article
language
英語
OAI-PMH Set
岡山大学
File Version
author
PubMed ID
DOI
Web of Science KeyUT
Related Url
isVersionOf https://doi.org/10.1080/09168451.2018.1459464
Funder Name
Ministry of Education, Culture, Sports, Science and Technology
助成番号
15K07841
15KK0282
17K0819709