Novel Catalytic Mechanism of Glycoside Hydrolysis Based on the Structure of an NAD+/Mn2+ -Dependent Phospho-Alpha-Glucosidase From Bacillus Subtilis

Structure. 2004 Sep;12(9):1619-29. doi: 10.1016/j.str.2004.06.020.

Abstract

GlvA, a 6-phospho-alpha-glucosidase from Bacillus subtilis, catalyzes the hydrolysis of maltose-6'-phosphate and belongs to glycoside hydrolase family GH4. GH4 enzymes are unique in their requirement for NAD(H) and a divalent metal for activity. We have determined the crystal structure of GlvA in complex with its ligands to 2.05 A resolution. Analyses of the active site architecture, in conjunction with mechanistic studies and precedent from the nucleotide diphosphate hexose dehydratases and other systems, suggest a novel mechanism of glycoside hydrolysis by GlvA that involves both the NAD(H) and the metal.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / enzymology*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Crystallography, X-Ray
  • Glycosides / metabolism*
  • L-Lactate Dehydrogenase / chemistry
  • L-Lactate Dehydrogenase / genetics
  • L-Lactate Dehydrogenase / metabolism
  • Manganese / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Molecular Structure
  • NAD / chemistry*
  • NAD / genetics
  • NAD / metabolism*
  • Protein Conformation*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Substrate Specificity
  • alpha-Glucosidases / chemistry*
  • alpha-Glucosidases / genetics
  • alpha-Glucosidases / metabolism

Substances

  • Bacterial Proteins
  • Glycosides
  • Recombinant Proteins
  • NAD
  • Manganese
  • L-Lactate Dehydrogenase
  • maltose-6'-phosphate glucosidase
  • alpha-Glucosidases

Associated data

  • PDB/1U8X