論文

査読有り
2011年9月

Specific Enzyme Complex of beta-1,4-Galactosyltransferase-II and Glucuronyltransferase-P Facilitates Biosynthesis of N-linked Human Natural Killer-1 (HNK-1) Carbohydrate

JOURNAL OF BIOLOGICAL CHEMISTRY
  • Tetsuya Kouno
  • ,
  • Yasuhiko Kizuka
  • ,
  • Naoki Nakagawa
  • ,
  • Toru Yoshihara
  • ,
  • Masahide Asano
  • ,
  • Shogo Oka

286
36
開始ページ
31337
終了ページ
31346
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1074/jbc.M111.233353
出版者・発行元
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Human natural killer-1 (HNK-1) carbohydrate is highly expressed in the nervous system and is involved in synaptic plasticity and dendritic spine maturation. This unique carbohydrate, consisting of a sulfated trisaccharide (HSO(3)-3GlcA beta 1-3Gal beta 1-4GlcNAc-), is biosynthesized by the successive actions of beta-1,4-galactosyltransferase (beta 4GalT), glucuronyltransferase (GlcAT-P and GlcAT-S), and sulfotransferase (HNK-1ST). A previous study showed that mice lacking beta 4GalT-II, one of seven beta 4GalTs, exhibited a dramatic loss of HNK-1 expression in the brain, although beta 4GalT-I-deficient mice did not. Here, we investigated the underlying molecular mechanism of the regulation of HNK-1 expression. First, focusing on a major HNK-1 carrier, neural cell adhesion molecule, we found that reduced expression of an N-linked HNK-1 carbohydrate caused by a deficiency of beta 4GalT-II is not likely due to a general loss of the beta 1,4-galactose residue as an acceptor for GlcAT-P. Instead, we demonstrated by co-immunoprecipitation and endoplasmic reticulum-retention analyses using Neuro2a (N2a) cells that beta 4GalT-II physically and specifically associates with GlcAT-P. In addition, we revealed by pull-down assay that Golgi luminal domains of beta 4GalT-II and GlcAT-P are sufficient for the complex to form. With an in vitro assay system, we produced the evidence that the kinetic efficiency k(cat)/K(m) of GlcAT-P in the presence of beta 4GalT-II was increased about 2.5-fold compared with that in the absence of beta 4GalT-II. Finally, we showed that co-expression of beta 4GalT-II and GlcAT-P increased HNK-1 expression on various glycoproteins in N2a cells, including neural cell adhesion molecule. These results indicate that the specific enzyme complex of beta 4GalT-II with GlcAT-P plays an important role in the biosynthesis of HNK-1 carbohydrate.

Web of Science ® 被引用回数 : 17

リンク情報
DOI
https://doi.org/10.1074/jbc.M111.233353
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/21771787
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000294487500029&DestApp=WOS_CPL
ID情報
  • DOI : 10.1074/jbc.M111.233353
  • ISSN : 0021-9258
  • PubMed ID : 21771787
  • Web of Science ID : WOS:000294487500029

エクスポート
BibTeX RIS