1H and 15N assignments and secondary structure of the starch-binding domain of glucoamylase from Aspergillus niger

Eur J Biochem. 1995 Oct 15;233(2):568-78. doi: 10.1111/j.1432-1033.1995.568_2.x.

Abstract

1H and 15N NMR resonance assignments of the granular starch-binding domain (SBD) of glucoamylase from Aspergillus niger have been made by multi-dimensional homonuclear and heteronuclear NMR techniques. Secondary structure analysis based on chemical shifts, 1H-1H NOEs, coupling constants and backbone amide exchange data indicates the presence of a well-defined beta-sheet structure. This consists of one parallel and five antiparallel pairs of beta-strands forming two beta-sheets. Cis-trans isomerisation of proline residues and O-glycosylation of threonine residues are observed and compared between the proteolytically derived SBD fragment and the recombinant protein. Structural features of the SBD in solution were compared to the X-ray crystal structure of a homologous domain of cyclodextrin glycosyltransferase from Bacillus circulans. There are some differences in the locations of the start and end of beta-strands but overall the two structures are very similar. This study will form the basis for the structure determination of the granular SBD and of its complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aspergillus niger / enzymology*
  • Binding Sites
  • Glucan 1,4-alpha-Glucosidase / chemistry*
  • Glycosylation
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Proline / chemistry
  • Protein Structure, Secondary*
  • Starch / metabolism*

Substances

  • Starch
  • Proline
  • Glucan 1,4-alpha-Glucosidase