4PEJ

Crystal structure of a computationally designed retro-aldolase, RA110.4 (Cys free)

Summary for 4PEJ

• Entry DOI: 10.2210/pdb4pej/pdb
• Related: 4PEK
• Descriptor: Retro-aldolase (2 entities in total)
• Functional Keywords: computationally designed enzyme, fluorescent probe, lyase
• Biological source: ARTIFICIAL GENE
• Total number of polymer chains: 2
• Total formula weight: 32001.91

Authors

Bhabha, G.,Zhang, X.,Liu, Y.,Ekiert, D.C. (deposition date: 2014-04-23, release date: 2015-04-08, Last modification date: 2023-09-27)

Primary citation

Liu, Y.,Zhang, X.,Tan, Y.L.,Bhabha, G.,Ekiert, D.C.,Kipnis, Y.,Bjelic, S.,Baker, D.,Kelly, J.W.
De novo-designed enzymes as small-molecule-regulated fluorescence imaging tags and fluorescent reporters.
J.Am.Chem.Soc., 136:13102-13105, 2014

PubMed Abstract: Enzyme-based tags attached to a protein-of-interest (POI) that react with a small molecule, rendering the conjugate fluorescent, are very useful for studying the POI in living cells. These tags are typically based on endogenous enzymes, so protein engineering is required to ensure that the small-molecule probe does not react with the endogenous enzyme in the cell of interest. Here we demonstrate that de novo-designed enzymes can be used as tags to attach to POIs. The inherent bioorthogonality of the de novo-designed enzyme-small-molecule probe reaction circumvents the need for protein engineering, since these enzyme activities are not present in living organisms. Herein, we transform a family of de novo-designed retroaldolases into variable-molecular-weight tags exhibiting fluorescence imaging, reporter, and electrophoresis applications that are regulated by tailored, reactive small-molecule fluorophores.

PubMed: 25209927
DOI: 10.1021/ja5056356

Experimental method

X-RAY DIFFRACTION (1.85 Å)