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標題: Determination of N-Glycosylation Site and Glycan Structures of Pectin Methylesterase in Jelly Fig (Ficus awkeotsang) Achenes
作者: Hsiao, E.S.L.
Chen, J.C.F.
Tsai, H.Y.
Khoo, K.H.
Chen, S.T.
Tzen, J.T.C.
關鍵字: Glycan structure;jelly fig;mass analyses;N-glycosylation;pectin;methylesterase;mosaic-virus movement;functional expression;purification;esterase;protein
Project: Journal of Agricultural and Food Chemistry
期刊/報告no:: Journal of Agricultural and Food Chemistry, Volume 57, Issue 15, Page(s) 6757-6763.
Pectin methylesterase (PME) in jelly fig (Ficus awkeotsang) achenes is an N-glycosylated enzyme responsible for the gelation of jelly curd. A recombinant jelly fig PME was overexpressed in Escherichia coli and confirmed by immunodetection and LC-nanoESI-MS/MS analysis. To identify the N-glycosylation site, native PME and its deglycosylated and recombinant forms, which lacked glycan, were purified and subjected to comparative MALDI-MS mapping of the corresponding tryptic fragments. The results showed that N-glycosylation occurred at Asn(153) of the mature jelly fig PME, the only glycosylation site predicted by its sequence analysis. The major N-glycans released from the native PME by PNGase IF were identified by MS/MS analyses as xylosylated, noncore fucosylated pauci-mannose, and complex-type structures. Molecular modeling of the 3D structure of jelly fig PME indicated that the N-glycan was putatively attached to the back region of the active site of this enzyme.
ISSN: 0021-8561
DOI: 10.1021/jf9011278
Appears in Collections:生物科技學研究所

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