ArticleProceedings of the National Academy of Sciences of the United States of America2002
Chemical complementation: a reaction-independent genetic assay for enzyme catalysis.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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24 citing papers in PubMed, 102 citations in OpenAlex.
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- Selecting Better Biocatalysts by Complementing Recoded Bacteria.Angewandte Chemie (International ed. in English) · 2023Article
- Evolution of a split RNA polymerase as a versatile biosensor platform.Nature chemical biology · 2017Article
- Studying disorders of vertebrate iron and heme metabolism using zebrafish.Methods in cell biology · 2017Review
- Methods for the directed evolution of proteins.Nature reviews. Genetics · 2015Review
- Transcriptional regulation improves the throughput of three-hybrid counter selections in Saccharomyces cerevisiae.Biotechnology journal · 2013Article
- Effective molarityIsrael journal of chemistry · 2013Article
- A Heritable Recombination system for synthetic Darwinian evolution in yeast.ACS synthetic biology · 2012Article
- Diversity in genetic in vivo methods for protein-protein interaction studies: from the yeast two-hybrid system to the mammalian split-luciferase system.Microbiology and molecular biology reviews : MMBR · 2012Review
- Applications of genetically-encoded biosensors for the construction and control of biosynthetic pathways.Metabolic engineering · 2012Review
- Identification and validation of protein targets of bioactive small molecules.Bioorganic & medicinal chemistry · 2012Review
- A human vitamin D receptor mutant activated by cholecalciferol.The Journal of steroid biochemistry and molecular biology · 2011Article
- Article
- The yeast three-hybrid system as an experimental platform to identify proteins interacting with small signaling molecules in plant cells: potential and limitations.Frontiers in plant science · 2011Article
- Nucleating the assembly of macromolecular complexes.Chembiochem : a European journal of chemical biology · 2010Article
- Synthesis of orthogonally reactive FK506 derivatives via olefin cross metathesis.Bioorganic & medicinal chemistry · 2009Article
- Evolution of efficient pathways for degradation of anthropogenic chemicals.Nature chemical biology · 2009Review
- High-throughput selection for cellulase catalysts using chemical complementation.Journal of the American Chemical Society · 2008Article
- Design and Applications of Bifunctional Small Molecules: Why Two Heads Are Better Than One.ACS chemical biology · 2008Review
- Identification of novel benzoylformate decarboxylases by growth selection.Applied and environmental microbiology · 2006Article
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7 authors at 1 institution in 1 country.
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Abstract
A high-throughput assay for enzyme activity has been developed that is reaction independent. In this assay, a small-molecule yeast three-hybrid system is used to link enzyme catalysis to transcription of a reporter gene in vivo. Here we demonstrate the feasibility of this approach by using a well-studied enzyme-catalyzed reaction, cephalosporin hydrolysis by the Enterobacter cloacae P99 cephalosporinase (beta-lactam hydrolase, EC ). We show that the three-hybrid system can be used to read out cephalosporinase activity in vivo as a change in the level of transcription of a lacZ reporter gene and that the wild-type cephalosporinase can be isolated from a pool of inactive mutants by using a lacZ screen. The assay has been designed so that it can be applied to different chemical reactions without changing the components of the three-hybrid system. A reaction-independent high-throughput assay for protein function should be a powerful tool for protein engineering and enzymology, drug discovery, and proteomics.
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