論文

査読有り
2006年12月

Molecular cloning and characterization of a novel human beta 1,3-glucosyltransferase, which is localized at the endoplasmic reticulum and glucosylates O-linked fucosylglycan on thrombospondin type 1 repeat domain

GLYCOBIOLOGY
  • Takashi Sato
  • Maiko Sato
  • Katsue Kiyohara
  • Maki Sogabe
  • Toshihide Shikanai
  • Norihiro Kikuchi
  • Akira Togayachi
  • Hiroyasu Ishida
  • Hiromi Ito
  • Akihiko Kameyama
  • Masanori Gotoh
  • Hisashi Narimatsu
  • 全て表示

16
12
開始ページ
1194
終了ページ
1206
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1093/glycob/cwl035
出版者・発行元
OXFORD UNIV PRESS INC

Protein O-linked fucosylation is an unusual glycosylation associated with many important biological functions such as Notch signaling. Two fucosylation pathways synthesizing O-fucosylglycans have been reported on cystein-knotted proteins, that is, on epidermal growth factor-like (EGF-like) domains and on thrombospondin Type 1 repeat (TSR) domains. We report here the molecular cloning and characterization of a novel beta 1,3-glucosyltransferase (beta 3Glc-T) that synthesizes a Glc beta 1,3Fuc alpha- structure on the TSR domain. We found a novel glycosyltransferase gene with beta 1,3-glycosyltransferase (beta 3GT) motifs in databases. The recombinant enzyme expressed in human embryonic kidney 293T (HEK293T) cells exhibited glucosyltransferase activity toward fucose-alpha-para-nitrophenyl (Fuc alpha-pNp). Thin-layer chromatography (TLC) analysis revealed that the product of the recombinant enzyme migrated to the same position as did the product of endogenous beta 3Glc-T of Chinese hamster ovary (CHO) cells. The two products could be digested by beta-glucosidase from almond and by exo-1,3-beta-glucanase from Trichoderma sp. These results strongly suggested that the product has the structure of Glc beta 1-3Fuc. Therefore, we named this novel enzyme beta 3Glc-T. Immunostaining revealed that FLAG-tagged beta 3Glc-T is an enzyme residing in the endoplasmic reticulum (ER) via retention signal, "REEL," which is a KDEL-like sequence, at the C-terminus. The TSR domain expressed in Escherichia coli was first fucosylated by the recombinant protein O-fucosyltransferase 2 (POFUT2), after which it became an acceptor substrate for the recombinant beta 3Glc-T, which could apparently transfer Glc to the fucosylated TSR domain. Our results suggest that a novel glycosyltransferase, beta 3Glc-T, contributes to the elongation of O-fucosylglycan and that this occurs specifically on TSR domains.

リンク情報
DOI
https://doi.org/10.1093/glycob/cwl035
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000242270800005&DestApp=WOS_CPL
ID情報
  • DOI : 10.1093/glycob/cwl035
  • ISSN : 0959-6658
  • eISSN : 1460-2423
  • Web of Science ID : WOS:000242270800005

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