3FCA

Genetic Incorporation of a Metal-ion Chelating Amino Acid into proteins as biophysical probe

Summary for 3FCA

• Entry DOI: 10.2210/pdb3fca/pdb
• Descriptor: Cysteine synthase, ZINC ION (3 entities in total)
• Functional Keywords: phasing, heavy metal, unnatural amino acid, metal binding, transferase
• Biological source: Thermus thermophilus
• Total number of polymer chains: 2
• Total formula weight: 62605.53

Authors

Wang, F.,Lee, H.,Spraggon, G.,Schultz, P.G. (deposition date: 2008-11-21, release date: 2009-02-17, Last modification date: 2024-03-27)

Primary citation

Lee, H.S.,Spraggon, G.,Schultz, P.G.,Wang, F.
Genetic incorporation of a metal-ion chelating amino acid into proteins as a biophysical probe.
J.Am.Chem.Soc., 131:2481-2483, 2009

PubMed Abstract: A metal-ion chelating amino acid, (8-hydroxyquinolin-3-yl)alanine, was genetically encoded in E. coli by an amber nonsense codon and corresponding orthogonal tRNA/aminoacyl-tRNA synthetase pair. The amino acid was incorporated into TM0665 protein, and the mutant protein was cocrystallized with Zn(2+) to determine the structure by SAD phasing. The structure showed a high occupancy of the heavy metal bound to the HQ-Ala residue, and the heavy metal provided excellent phasing power to determine the structure. This method also facilitates the de novo design of metalloproteins with novel structures and functions, including fluorescent sensors.

PubMed: 19193005
DOI: 10.1021/ja808340b

Experimental method

X-RAY DIFFRACTION (2.149 Å)