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

Loading...
Thumbnail Image

Date

2021-01-15

Authors

Gamiz-Arco, Gloria
Gutierrez-Rus, Luis I
Risso, Valeria A
Ibarra-Molero, Beatriz
Hoshino, Yosuke
Petrović, Dušan
Justicia, Jose
Cuerva, Juan Manuel
Romero-Rivera, Adrian
Seelig, Burckhard

Advisors

Journal Title

Journal ISSN

Volume Title

Publisher

Metrics
Google Scholar
Export

Research Projects

Organizational Units

Journal Issue

Abstract

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.

Description

MeSH Terms

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

DeCS Terms

CIE Terms

Keywords

Citation