https://scholars.lib.ntu.edu.tw/handle/123456789/143901
標題: | Identification of Further Elongation and Branching of Dimeric Type 1 Chain on Lactosylceramides from Colonic Adenocarcinoma by Tandem Mass Spectrometry Sequencing Analyses | 作者: | Fan, Yao-Yun Yu, Shin-Yi Ito, Hiromi Kameyama, Akihiko Sato, Takashi Lin, Chi-Hung Yu, Lung-Chih Narimatsu, Hisashi Khoo, Kay-Hooi |
公開日期: | 2008 | 起(迄)頁: | 16455-16468 | 來源出版物: | Journal of Biological Chemistry | 摘要: | Mammalian glycan chain elongation is mostly based on extending the type 2 chain, Galβ1-4GlcNAc, whereas the corresponding type 1 chain, Galβ1-3GlcNAc, is not normally extended. In a broader context of developing high sensitivity mass spectrometry methodologies for glycomic identification of Lea versus Lex and linear versus branched poly-N-acetyllactosamine (polyLacNAc), we have now shown that the dimeric type 1 glycan chain, as carried on the lactosylceramides of a human colonic adenocarcinoma cell line, Colo205, not only can be further extended linearly but can likewise be branched at C6 of 3-linked Gal in a manner similar to polyLacNAc. A combination of chemical and enzymatic derivatization coupled with advanced mass spectrometry analyses afforded unambiguous identification of a complex mixture of type 1 and 2 hybrids as well as those fucosylated variants founded exclusively on linear and branched trimeric type 1 chain. We further showed by in vitro enzymatic synthesis that extended type 1 and the hybrid chains can be branched by all three forms of the human I branching enzymes (IGnT) currently identified but with lower efficiency and stringency with respect to branching site preference. Importantly, it was found that a better substrate is one that carries a Gal site for branching that is extended at the non-reducing end by a type 2 and not a type 1 unit, whereas the IGnTs are less discriminative with respect to whether the targeted Gal site is itself β3- or β4-linked to GlcNAc at the reducing end. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/163236 https://www.scopus.com/inward/record.uri?eid=2-s2.0-47749108740&doi=10.1074%2fjbc.M707274200&partnerID=40&md5=a5caa05eb87495465daa7a83202f86fa |
ISSN: | 00219258 | DOI: | 10.1074/jbc.M707274200 | SDG/關鍵字: | Cell culture; Electric field effects; Mammals; Mass spectrometers; Mass spectrometry; Spectrometers; Spectrometry; Spectrum analysis; Adenocarcinoma cell lines; Branched polies; Branching enzymes; Complex mixtures; Derivatization; Enzymatic syntheses; Glycan chains; High sensitivities; Hybrid chains; In vitro; Mass spectrometry analysis; Reducing ends; Sequencing analysis; Site preferences; Tandem mass spectrometries; High performance liquid chromatography; 1,4 alpha glucan branching enzyme; aminosugar; glycan; lactosylceramide; poly n acteyllactosamine; unclassified drug; fucose; lactosylceramide; messenger RNA; poly n acetyllactosamine; poly-N-acetyllactosamine; polysaccharide; article; colon adenocarcinoma; controlled study; derivatization; enzyme synthesis; fucosylation; human; human cell; hybrid; matrix assisted laser desorption ionization time of flight mass spectrometry; outcome assessment; priority journal; sequence analysis; tandem mass spectrometry; adenocarcinoma; cell fractionation; chemistry; colon tumor; dimerization; exon; mass spectrometry; metabolism; methodology; tumor cell line; Mammalia; Adenocarcinoma; Cell Line, Tumor; Colonic Neoplasms; Dimerization; Exons; Fucose; Humans; Lactosylceramides; Mass Spectrometry; Polysaccharides; RNA, Messenger; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Subcellular Fractions; Tandem Mass Spectrometry |
顯示於: | 生化科學研究所 |
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