Identification of the catalytic acid-base residue of arthrobacter endo-β-N-acetylglucosaminidase by chemical rescue of an inactive mutant

Kiyotaka Fujita, Reiko Sato, Kazunori Toma, Kanefumi Kitahara, Toshihiko Suganuma, Kenji Yamamoto, Kaoru Takegawa

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Arthrobacter endo-β-N-acetylglucosaminidase (Endo-A), a member of glycoside hydrolase (GH) family 85, catalyses the hydrolysis and transglycosylation of asparagine-linked oligosaccharides of glycoproteins with retention of anomeric configuration. Glu-173 of Endo-A is a catalytically essential amino acid residue, and the corresponding residue is conserved in all GH family 85 members. The catalytic activity of Endo-A E173A mutant was rescued by the addition of sodium azide or sodium formate. Furthermore, the produced β-glycosyl azide (Man 5GlcNAc-β-N 3) retained the anomeric configuration, indicating that Glu-173 is the catalytic acid-base residue of Endo-A. This is the first identification of the catalytic residue for GH family 85 endo-β-N-acetylglucosaminidases.

Original languageEnglish
Pages (from-to)301-306
Number of pages6
JournalJournal of biochemistry
Volume142
Issue number3
DOIs
Publication statusPublished - Sep 1 2007

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Arthrobacter
Acetylglucosaminidase
Glycoside Hydrolases
formic acid
Acids
Sodium Azide
Essential Amino Acids
Azides
Asparagine
Oligosaccharides
Hydrolysis
Catalyst activity
Glycoproteins
endo-alpha-sialidase

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Identification of the catalytic acid-base residue of arthrobacter endo-β-N-acetylglucosaminidase by chemical rescue of an inactive mutant. / Fujita, Kiyotaka; Sato, Reiko; Toma, Kazunori; Kitahara, Kanefumi; Suganuma, Toshihiko; Yamamoto, Kenji; Takegawa, Kaoru.

In: Journal of biochemistry, Vol. 142, No. 3, 01.09.2007, p. 301-306.

Research output: Contribution to journalArticle

Fujita, Kiyotaka ; Sato, Reiko ; Toma, Kazunori ; Kitahara, Kanefumi ; Suganuma, Toshihiko ; Yamamoto, Kenji ; Takegawa, Kaoru. / Identification of the catalytic acid-base residue of arthrobacter endo-β-N-acetylglucosaminidase by chemical rescue of an inactive mutant. In: Journal of biochemistry. 2007 ; Vol. 142, No. 3. pp. 301-306.
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AU - Toma, Kazunori

AU - Kitahara, Kanefumi

AU - Suganuma, Toshihiko

AU - Yamamoto, Kenji

AU - Takegawa, Kaoru

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