Kinetics of iterative carbohydrate transfer to polysaccharide catalyzed by chondroitin polymerase on a highly sensitive flow-type 27 MHz quartz-crystal microbalance

Toshiaki Mori, Takayuki Kodera, Hiroshi Yoshimine, Yoshimitsu Kakuta, Nobuo Sugiura, Koji Kimata, Yoshio Okahata

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

Using a highly sensitive flow-type 27 MHz quartz crystal microbalance, we could detect a small mass change during stepwise and alternating one-sugar transfer of glucuronic acid (GlcA) and N-acetylgalactosamine (GalNAc) to an acceptor, catalyzed by chondroitin polymerase from Escherichia coli strain K4 (K4CP), and analyze the elongation mechanism of K4CP. K4CP was found to bind strongly to a chondroitin acceptor (K d=0.97 μM). Although the binding affinity and the catalytic rate constant for each monomer were considerably different, the apparent catalytic efficiency (k cat/ K m) was similar (6.3×10 4 M -1 s -1 for GlcA transfer and 3.4×10 4 M -1 s -1 for the GalNAc transfer). This is reasonable for the smooth alternating elongation of GlcA and GalNAc on the acceptor. This is the first study to report the determination of kinetic parameters for enzymatic, alternated, sugar elongation. One-sugar transfer: Kinetics of one-sugar transfer can be evaluated by using a highly sensitive flow-type quartz-crystal microbalance. Binding affinity and turnover number for UDP-GlcA and UDP-GalNAc are quite different. The catalytic efficiency is similar for GlcA transfer and GalNAc transfer (see scheme).

元の言語英語
ページ(範囲)7389-7393
ページ数5
ジャーナルChemistry - A European Journal
18
発行部数24
DOI
出版物ステータス出版済み - 6 11 2012

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Chondroitin
Glucuronic Acid
Quartz crystal microbalances
Polysaccharides
Carbohydrates
Sugars
Elongation
Kinetics
Acids
Uridine Diphosphate Glucuronic Acid
Acetylgalactosamine
Uridine Diphosphate
Kinetic parameters
Escherichia coli
Rate constants
Monomers

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

これを引用

Kinetics of iterative carbohydrate transfer to polysaccharide catalyzed by chondroitin polymerase on a highly sensitive flow-type 27 MHz quartz-crystal microbalance. / Mori, Toshiaki; Kodera, Takayuki; Yoshimine, Hiroshi; Kakuta, Yoshimitsu; Sugiura, Nobuo; Kimata, Koji; Okahata, Yoshio.

:: Chemistry - A European Journal, 巻 18, 番号 24, 11.06.2012, p. 7389-7393.

研究成果: ジャーナルへの寄稿記事

Mori, Toshiaki ; Kodera, Takayuki ; Yoshimine, Hiroshi ; Kakuta, Yoshimitsu ; Sugiura, Nobuo ; Kimata, Koji ; Okahata, Yoshio. / Kinetics of iterative carbohydrate transfer to polysaccharide catalyzed by chondroitin polymerase on a highly sensitive flow-type 27 MHz quartz-crystal microbalance. :: Chemistry - A European Journal. 2012 ; 巻 18, 番号 24. pp. 7389-7393.
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