Crystals in Minutes: Instant On-Site Microcrystallisation of Various Flavours of the CYP102A1 (P450BM3) Haem Domain

Angew Chem Int Ed Engl. 2020 May 4;59(19):7611-7618. doi: 10.1002/anie.201913407. Epub 2020 Mar 11.


Despite CYP102A1 (P450BM3) representing one of the most extensively researched metalloenzymes, crystallisation of its haem domain upon modification can be a challenge. Crystal structures are indispensable for the efficient structure-based design of P450BM3 as a biocatalyst. The abietane diterpenoid derivative N-abietoyl-l-tryptophan (AbiATrp) is an outstanding crystallisation accelerator for the wild-type P450BM3 haem domain, with visible crystals forming within 2 hours and diffracting to a near-atomic resolution of 1.22 Å. Using these crystals as seeds in a cross-microseeding approach, an assortment of P450BM3 haem domain crystal structures, containing previously uncrystallisable decoy molecules and diverse artificial metalloporphyrins binding various ligand molecules, as well as heavily tagged haem-domain variants, could be determined. Some of the structures reported herein could be used as models of different stages of the P450BM3 catalytic cycle.

Keywords: crystal growth; cytochromes; decoy molecules; enzyme models; protein structures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus megaterium / chemistry
  • Bacterial Proteins / chemistry*
  • Catalysis
  • Crystallization / methods*
  • Cytochrome P-450 Enzyme System / chemistry*
  • Heme / chemistry
  • Indicators and Reagents
  • Metalloporphyrins / chemical synthesis
  • Mutagenesis, Site-Directed
  • NADPH-Ferrihemoprotein Reductase / chemistry*
  • Protein Binding
  • Substrate Specificity
  • X-Ray Diffraction


  • Bacterial Proteins
  • Indicators and Reagents
  • Metalloporphyrins
  • Heme
  • Cytochrome P-450 Enzyme System
  • NADPH-Ferrihemoprotein Reductase
  • flavocytochrome P450 BM3 monoxygenases