An artificial metalloprotein with metal-adaptive coordination sites and Ni-dependent quercetinase activity

Morane Beaumet, Anica Dose, Alois Bräuer, Jean Pierre Mahy, Wadih Ghattas, Michael Groll, Corinna R. Hess

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Engineering non-native metal active sites into proteins using canonical amino acids offers many advantages but is hampered by significant challenges. The TIM barrel protein, imidazole glycerol phosphate synthase from the hyperthermophilic organism Thermotoga maritima (tHisF), is well-suited for the construction of artificial metalloenzymes by this approach. To this end, we have generated a tHisF variant (tHisFEHH) with a Glu/His/His motif for metal ion coordination. Crystal structures of ZnII:tHisFEHH and NiII:tHisFEHH reveal that both metal ions bind to the engineered histidines. However, the two metals bind at distinct sites with different geometries, demonstrating the adaptability of tHisF. Only ZnII additionally ligates the Glu residue and adopts a tetrahedral geometry. The pseudo-octahedral NiII site comprises the two His and a native Ser residue. NiII:tHisFEHH catalyzes the oxidative cleavage of the flavanols quercetin and myricetin, providing an unprecedented example of an artificial metalloprotein with quercetinase activity.

Original languageEnglish
Article number111914
JournalJournal of Inorganic Biochemistry
Volume235
DOIs
StatePublished - Oct 2022

Keywords

  • Artificial enzyme
  • Metalloprotein
  • Nickel
  • Quercetinase
  • Zinc
  • tHisF

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