Structural Basis of the γ-Lactone-Ring Formation in Ascorbic Acid Biosynthesis by the Senescence Marker Protein-30/Gluconolactonase

Shingo Aizawa, Miki Senda, Ayaka Harada, Naoki Maruyama, Tetsuo Ishida, Toshiro Aigaki, Akihito Ishigami, Toshiya Senda

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The senescence marker protein-30 (SMP30), which is also called regucalcin, exhibits gluconolactonase (GNL) activity. Biochemical and biological analyses revealed that SMP30/GNL catalyzes formation of the γ-lactone-ring of l-gulonate in the ascorbic acid biosynthesis pathway. The molecular basis of the γ-lactone formation, however, remains elusive due to the lack of structural information on SMP30/GNL in complex with its substrate. Here, we report the crystal structures of mouse SMP30/GNL and its complex with xylitol, a substrate analogue, and those with 1,5-anhydro-d-glucitol and d-glucose, product analogues. Comparison of the crystal structure of mouse SMP30/GNL with other related enzymes has revealed unique characteristics of mouse SMP30/GNL. First, the substrate-binding pocket of mouse SMP30/GNL is designed to specifically recognize monosaccharide molecules. The divalent metal ion in the active site and polar residues lining the substrate-binding cavity interact with hydroxyl groups of substrate/product analogues. Second, in mouse SMP30/GNL, a lid loop covering the substrate-binding cavity seems to hamper the binding of l-gulonate in an extended (or all-trans) conformation; l-gulonate seems to bind to the active site in a folded conformation. In contrast, the substrate-binding cavities of the other related enzymes are open to the solvent and do not have a cover. This structural feature of mouse SMP30/GNL seems to facilitate the γ-lactone-ring formation.

Original languageEnglish
Article numbere53706
JournalPloS one
Volume8
Issue number1
DOIs
Publication statusPublished - 2013 Jan 29
Externally publishedYes

Fingerprint

gluconolactonase
Biosynthesis
Lactones
lactones
Ascorbic Acid
ascorbic acid
biosynthesis
Substrates
Proteins
proteins
mice
crystal structure
active sites
Conformations
Catalytic Domain
Crystal structure
Perognathus
Xylitol
xylitol
lids

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Structural Basis of the γ-Lactone-Ring Formation in Ascorbic Acid Biosynthesis by the Senescence Marker Protein-30/Gluconolactonase. / Aizawa, Shingo; Senda, Miki; Harada, Ayaka; Maruyama, Naoki; Ishida, Tetsuo; Aigaki, Toshiro; Ishigami, Akihito; Senda, Toshiya.

In: PloS one, Vol. 8, No. 1, e53706, 29.01.2013.

Research output: Contribution to journalArticle

Aizawa, Shingo ; Senda, Miki ; Harada, Ayaka ; Maruyama, Naoki ; Ishida, Tetsuo ; Aigaki, Toshiro ; Ishigami, Akihito ; Senda, Toshiya. / Structural Basis of the γ-Lactone-Ring Formation in Ascorbic Acid Biosynthesis by the Senescence Marker Protein-30/Gluconolactonase. In: PloS one. 2013 ; Vol. 8, No. 1.
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