5 publications

5 publications

A Designed Supramolecular Protein Assembly with In Vivo Enzymatic Activity

Tezcan, F. A.

Science, 2014, 10.1126/science.1259680

The generation of new enzymatic activities has mainly relied on repurposing the interiors of preexisting protein folds because of the challenge in designing functional, three-dimensional protein structures from first principles. Here we report an artificial metallo-β-lactamase, constructed via the self-assembly of a structurally and functionally unrelated, monomeric redox protein into a tetrameric assembly that possesses catalytic zinc sites in its interfaces. The designed metallo-β-lactamase is functional in the Escherichia coli periplasm and enables the bacteria to survive treatment with ampicillin. In vivo screening of libraries has yielded a variant that displays a catalytic proficiency [(kcat/Km)/kuncat] for ampicillin hydrolysis of 2.3 × 106 and features the emergence of a highly mobile loop near the active site, a key component of natural β-lactamases to enable substrate interactions.


Metal: Zn
Ligand type: Amino acid
Host protein: Cytochrome cb562
Anchoring strategy: Dative
Optimization: Genetic
Max TON: ---
ee: ---
PDB: 4U9E
Notes: ---

Design and Construction of Functional Supramolecular Metalloprotein Assemblies

Review

Tezcan, F. A.

Acc. Chem. Res., 2019, 10.1021/acs.accounts.8b00617


Notes: ---

Importance of Scaffold Flexibility/Rigidity in the Design and Directed Evolution of Artificial Metallo-β-Lactamases

Song, W. J.; Tezcan, F. A.

J. Am. Chem. Soc., 2017, 10.1021/jacs.7b08981


Metal: Zn
Ligand type: Amino acid
Host protein: Zn8:AB54
Anchoring strategy: Dative
Optimization: Genetic
Reaction: Hydrolysis
Max TON: ---
ee: ---
PDB: 5XZI
Notes: Supramolecular protein scaffold constructed from cytochrome cb562 building blocks, Ampicillin hydrolysis: kcat/KM = 130 min-1 * M-1

Metal: Zn
Ligand type: Amino acid
Host protein: Zn8:AB54 (mutant C96T)
Anchoring strategy: Dative
Optimization: Genetic
Reaction: Hydrolysis
Max TON: ---
ee: ---
PDB: 5XZJ
Notes: Supramolecular protein scaffold constructed from cytochrome cb562 building blocks, Ampicillin hydrolysis: kcat/KM = 210 min-1 * M-1

Interfacial Metal Coordination in Engineered Protein and Peptide Assemblies

Review

Tezcan, F. A.

Curr. Opin. Chem. Biol., 2014, 10.1016/j.cbpa.2013.12.013


Notes: ---

Metal-Directed Design of Supramolecular Protein Assemblies

Review

Tezcan, F. A.

Methods Enzymol., 2016, 10.1016/bs.mie.2016.05.009


Notes: ---