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Photo Lynn Kamerlin

Lynn Kamerlin

Professor

Photo Lynn Kamerlin

Heme-binding enables allosteric modulation in an ancient TIM-barrel glycosidase

Author

  • Gloria Gamiz-Arco
  • Luis I Gutierrez-Rus
  • Valeria A Risso
  • Beatriz Ibarra-Molero
  • Yosuke Hoshino
  • Dušan Petrović
  • Jose Justicia
  • Juan Manuel Cuerva
  • Adrian Romero-Rivera
  • Burckhard Seelig
  • Jose A Gavira
  • Shina C L Kamerlin
  • Eric A Gaucher
  • Jose M Sanchez-Ruiz
Show all

Summary, in English

Glycosidases are phylogenetically widely distributed enzymes that are crucial for the cleavage of glycosidic bonds. Here, we present the exceptional properties of a putative ancestor of bacterial and eukaryotic family-1 glycosidases. The ancestral protein shares the TIM-barrel fold with its modern descendants but displays large regions with greatly enhanced conformational flexibility. Yet, the barrel core remains comparatively rigid and the ancestral glycosidase activity is stable, with an optimum temperature within the experimental range for thermophilic family-1 glycosidases. None of the ∼5500 reported crystallographic structures of ∼1400 modern glycosidases show a bound porphyrin. Remarkably, the ancestral glycosidase binds heme tightly and stoichiometrically at a well-defined buried site. Heme binding rigidifies this TIM-barrel and allosterically enhances catalysis. Our work demonstrates the capability of ancestral protein reconstructions to reveal valuable but unexpected biomolecular features when sampling distant sequence space. The potential of the ancestral glycosidase as a scaffold for custom catalysis and biosensor engineering is discussed.

Publishing year

2021-01-15

Language

English

Publication/Series

Nature Communications

Volume

12

Issue

1

Document type

Journal article

Publisher

Nature Publishing Group

Keywords

  • Allosteric Regulation
  • Amino Acid Sequence/genetics
  • Bacteria/enzymology
  • Crystallography, X-Ray
  • Eukaryota/enzymology
  • Glycoside Hydrolases/genetics
  • Heme/metabolism
  • Molecular Dynamics Simulation
  • Phylogeny
  • Protein Structure, Secondary
  • Sequence Homology, Amino Acid

Status

Published

ISBN/ISSN/Other

  • ISSN: 2041-1723