5D3U

Crystal structure of the 5-selective H176F mutant of Cytochrome TxtE

Summary for 5D3U

• Entry DOI: 10.2210/pdb5d3u/pdb
• Related: 5D40
• Descriptor: P450-like protein, PROTOPORPHYRIN IX CONTAINING FE, TRYPTOPHAN, ... (6 entities in total)
• Functional Keywords: cytochrome, p450, heme, regioselectivity, f/g loop, oxidoreductase
• Biological source: Streptomyces scabies (strain 87.22)
• Total number of polymer chains: 2
• Total formula weight: 96150.06

Authors

Cahn, J.K.B.,Dodani, S.C.,Arnold, F.H. (deposition date: 2015-08-06, release date: 2016-06-22, Last modification date: 2023-09-27)

Primary citation

Dodani, S.C.,Kiss, G.,Cahn, J.K.,Su, Y.,Pande, V.S.,Arnold, F.H.
Discovery of a regioselectivity switch in nitrating P450s guided by molecular dynamics simulations and Markov models.
Nat.Chem., 8:419-425, 2016

PubMed Abstract: The dynamic motions of protein structural elements, particularly flexible loops, are intimately linked with diverse aspects of enzyme catalysis. Engineering of these loop regions can alter protein stability, substrate binding and even dramatically impact enzyme function. When these flexible regions are unresolvable structurally, computational reconstruction in combination with large-scale molecular dynamics simulations can be used to guide the engineering strategy. Here we present a collaborative approach that consists of both experiment and computation and led to the discovery of a single mutation in the F/G loop of the nitrating cytochrome P450 TxtE that simultaneously controls loop dynamics and completely shifts the enzyme's regioselectivity from the C4 to the C5 position of L-tryptophan. Furthermore, we find that this loop mutation is naturally present in a subset of homologous nitrating P450s and confirm that these uncharacterized enzymes exclusively produce 5-nitro-L-tryptophan, a previously unknown biosynthetic intermediate.

PubMed: 27102675
DOI: 10.1038/nchem.2474

Experimental method

X-RAY DIFFRACTION (1.45 Å)