8HJX

Crystal structure of a cupin protein (tm1459, H52A/H58E mutant) in copper (Cu) substituted form

Summary for 8HJX

• Entry DOI: 10.2210/pdb8hjx/pdb
• Descriptor: Cupin_2 domain-containing protein, COPPER (II) ION (3 entities in total)
• Functional Keywords: artificial metalloenzymes, non-heme copper enzyme, cupin, metal binding protein
• Biological source: Thermotoga maritima MSB8
• Total number of polymer chains: 2
• Total formula weight: 26957.17

Authors

Matsumoto, R.,Kurisu, G.,Fujieda, N. (deposition date: 2022-11-24, release date: 2023-06-14, Last modification date: 2024-11-06)

Primary citation

Matsumoto, R.,Yoshioka, S.,Yuasa, M.,Morita, Y.,Kurisu, G.,Fujieda, N.
An artificial metallolyase with pliable 2-His-1-carboxylate facial triad for stereoselective Michael addition.
Chem Sci, 14:3932-3937, 2023

PubMed Abstract: We repurposed the metal-binding site of a cupin superfamily protein into the 2-His-1-carboxylate facial triad, which is one of the common motifs in natural non-heme enzymes, to construct artificial metalloenzymes that can catalyze new-to-nature reactions. The Cu-H52A/H58E variant catalyzed the stereoselective Michael addition reaction and was found to bear a flexible metal-binding site in the high-resolution crystal structure. Furthermore, the H52A/H58E/F104W mutant accommodated a water molecule, which was supported by Glu58 and Trp104 residues hydrogen bonding, presumably leading to high stereoselectivity. Thus, the 2-His-1-carboxylate facial triad was confirmed to be a versatile and promising metal-binding motif for abiological and canonical biological reactions.

PubMed: 37035687
DOI: 10.1039/d2sc06809e

Experimental method

X-RAY DIFFRACTION (1.15 Å)