Structural insights into the difference in substrate recognition of two mannoside phosphorylases from two GH130 subfamilies

Yuxin Ye, Wataru Saburi, Rei Odaka, Koji Kato, Naofumi Sakurai, Keisuke Komoda, Mamoru Nishimoto, Motomitsu Kitaoka, Haruhide Mori, Min Yao

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In Ruminococcus albus, 4-O-β-d-mannosyl-d-glucose phosphorylase (RaMP1) and β-(1,4)-mannooligosaccharide phosphorylase (RaMP2) belong to two subfamilies of glycoside hydrolase family 130. The two enzymes phosphorolyze β-mannosidic linkages at the nonreducing ends of their substrates, and have substantially diverse substrate specificity. The differences in their mechanism of substrate binding have not yet been fully clarified. In the present study, we report the crystal structures of RaMP1 with/without 4-O-β-d-mannosyl-d-glucose and RaMP2 with/without β-(1→4)-mannobiose. The structures of the two enzymes differ at the +1 subsite of the substrate-binding pocket. Three loops are proposed to determine the different substrate specificities. One of these loops is contributed from the adjacent molecule of the oligomer structure. In RaMP1, His245 of loop 3 forms a hydrogen-bond network with the substrate through a water molecule, and is indispensible for substrate binding.

Original languageEnglish
Pages (from-to)828-837
Number of pages10
JournalFEBS Letters
Volume590
Issue number6
DOIs
Publication statusPublished - Mar 1 2016
Externally publishedYes

Keywords

  • 4-O-β- d -mannosyl- d -glucose phosphorylase
  • glycoside hydro-lase family 130
  • hydrogen bond-network
  • substrate recognition
  • X-ray crystallography
  • β-1,4-mannooligosaccharide phosphorylase

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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  • Cite this

    Ye, Y., Saburi, W., Odaka, R., Kato, K., Sakurai, N., Komoda, K., Nishimoto, M., Kitaoka, M., Mori, H., & Yao, M. (2016). Structural insights into the difference in substrate recognition of two mannoside phosphorylases from two GH130 subfamilies. FEBS Letters, 590(6), 828-837. https://doi.org/10.1002/1873-3468.12105