Crystal structure of the covalent intermediate of human cytosolic β-glucosidase

Junji Noguchi, Yasuhiro Hayashi, Yuichi Baba, Nozomu Okino, Makoto Kimura, Makoto Ito, Yoshimitsu Kakuta

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Human cytosolic β-glucosidase, also known as klotho-related protein (KLrP, GBA3), is an enzyme that hydrolyzes various β-d-glucosides, including glucosylceramide. We recently reported the crystal structure of KLrP in complex with glucose [Y. Hayashi, N. Okino, Y. Kakuta, T. Shikanai, M. Tani, H. Narimatsu, M. Ito, Klotho-related protein is a novel cytosolic neutral beta-glycosylceramidase, J. Biol. Chem. 282 (2007) 30889-30900]. Here, we report the crystal structure of a covalent intermediate of the KLrP mutant E165Q, in which glucose was covalently bound to a nucleophile, Glu373. The structure confirms the double displacement mechanism of the retaining β-glucosidase. In addition, the structure suggests that a water molecule could be involved in the stabilization of transition states through a sugar, 2-hydroxyl.

Original languageEnglish
Pages (from-to)549-552
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume374
Issue number3
DOIs
Publication statusPublished - Sep 26 2008

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Glucosidases
Lactase-Phlorizin Hydrolase
Crystal structure
Glucosylceramides
Glucose
Nucleophiles
Glucosides
Sugars
Hydroxyl Radical
Proteins
Stabilization
Molecules
Water
Enzymes
klotho protein

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Crystal structure of the covalent intermediate of human cytosolic β-glucosidase. / Noguchi, Junji; Hayashi, Yasuhiro; Baba, Yuichi; Okino, Nozomu; Kimura, Makoto; Ito, Makoto; Kakuta, Yoshimitsu.

In: Biochemical and Biophysical Research Communications, Vol. 374, No. 3, 26.09.2008, p. 549-552.

Research output: Contribution to journalArticle

Noguchi, Junji ; Hayashi, Yasuhiro ; Baba, Yuichi ; Okino, Nozomu ; Kimura, Makoto ; Ito, Makoto ; Kakuta, Yoshimitsu. / Crystal structure of the covalent intermediate of human cytosolic β-glucosidase. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 374, No. 3. pp. 549-552.
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AU - Ito, Makoto

AU - Kakuta, Yoshimitsu

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AB - Human cytosolic β-glucosidase, also known as klotho-related protein (KLrP, GBA3), is an enzyme that hydrolyzes various β-d-glucosides, including glucosylceramide. We recently reported the crystal structure of KLrP in complex with glucose [Y. Hayashi, N. Okino, Y. Kakuta, T. Shikanai, M. Tani, H. Narimatsu, M. Ito, Klotho-related protein is a novel cytosolic neutral beta-glycosylceramidase, J. Biol. Chem. 282 (2007) 30889-30900]. Here, we report the crystal structure of a covalent intermediate of the KLrP mutant E165Q, in which glucose was covalently bound to a nucleophile, Glu373. The structure confirms the double displacement mechanism of the retaining β-glucosidase. In addition, the structure suggests that a water molecule could be involved in the stabilization of transition states through a sugar, 2-hydroxyl.

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