7U6H

HalD with ornithine and alpha-ketoglutarate

Summary for 7U6H

• Entry DOI: 10.2210/pdb7u6h/pdb
• Descriptor: Halogenase D, NICKEL (II) ION, CHLORIDE ION, ... (9 entities in total)
• Functional Keywords: biosynthetic protein
• Biological source: Pseudomonas kilonensis
• Total number of polymer chains: 4
• Total formula weight: 126281.48

Authors

Swenson, C.V.,Neugebauer, M.E.,Kissman, E.N.,Chang, M.C.Y. (deposition date: 2022-03-04, release date: 2023-03-15, Last modification date: 2023-10-25)

Primary citation

Kissman, E.N.,Neugebauer, M.E.,Sumida, K.H.,Swenson, C.V.,Sambold, N.A.,Marchand, J.A.,Millar, D.C.,Chang, M.C.Y.
Biocatalytic control of site-selectivity and chain length-selectivity in radical amino acid halogenases.
Proc.Natl.Acad.Sci.USA, 120:e2214512120-e2214512120, 2023

PubMed Abstract: Biocatalytic C-H activation has the potential to merge enzymatic and synthetic strategies for bond formation. Fe/αKG-dependent halogenases are particularly distinguished for their ability both to control selective C-H activation as well as to direct group transfer of a bound anion along a reaction axis separate from oxygen rebound, enabling the development of new transformations. In this context, we elucidate the basis for the selectivity of enzymes that perform selective halogenation to yield 4-Cl-lysine (BesD), 5-Cl-lysine (HalB), and 4-Cl-ornithine (HalD), allowing us to probe how site-selectivity and chain length selectivity are achieved. We now report the crystal structure of the HalB and HalD, revealing the key role of the substrate-binding lid in positioning the substrate for C vs C chlorination and recognition of lysine vs ornithine. Targeted engineering of the substrate-binding lid further demonstrates that these selectivities can be altered or switched, showcasing the potential to develop halogenases for biocatalytic applications.

PubMed: 36913566
DOI: 10.1073/pnas.2214512120

Experimental method

X-RAY DIFFRACTION (2 Å)