6K9S

Structure of the Carbonylruthenium Mesoporphyrin IX-Reconstituted CYP102A1 Haem Domain with N-Abietoyl-L-Tryptophan

Summary for 6K9S

• Entry DOI: 10.2210/pdb6k9s/pdb
• Descriptor: Bifunctional cytochrome P450/NADPH--P450 reductase, [3,3'-(7,12-diethyl-3,8,13,17-tetramethylporphyrin-2,18-diyl-kappa~4~N~21~,N~22~,N~23~,N~24~)dipropanoato(2-)]ruthenium, CARBON MONOXIDE, ... (6 entities in total)
• Functional Keywords: monooxygenase, oxidoreductase
• Biological source: Bacillus megaterium
• Total number of polymer chains: 2
• Total formula weight: 108163.37

Authors

Stanfield, J.K.,Omura, K.,Kasai, C.,Sugimoto, H.,Shiro, Y.,Watanabe, Y.,Shoji, O. (deposition date: 2019-06-17, release date: 2020-03-18, Last modification date: 2023-11-22)

Primary citation

Stanfield, J.K.,Omura, K.,Matsumoto, A.,Kasai, C.,Sugimoto, H.,Shiro, Y.,Watanabe, Y.,Shoji, O.
Crystals in Minutes: Instant On-Site Microcrystallisation of Various Flavours of the CYP102A1 (P450BM3) Haem Domain.
Angew.Chem.Int.Ed.Engl., 59:7611-7618, 2020

PubMed Abstract: 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.

PubMed: 32157795
DOI: 10.1002/anie.201913407

Experimental method

X-RAY DIFFRACTION (1.55 Å)