Mutagenesis and selection of PDZ domains that bind new protein targets
| Autor: | Christopher M. Hovens, Bruno Catimel, Thomas Baechi, Karin Moelling, Stefan Schneider, Michael Buchert, Steven A. Stacker, Oleg Georgiev, Michael M. Halford |
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| Rok vydání: | 1999 |
| Předmět: |
PDZ domain
Biomedical Engineering Mutagenesis (molecular biology technique) Bioengineering Plasma protein binding Biology Applied Microbiology and Biotechnology Cell Line Mice Animals Humans Amino Acid Sequence Binding site Peptide sequence DNA Primers Base Sequence Sequence Homology Amino Acid Proteins Protein engineering Subcellular localization Transmembrane protein Cell biology Biochemistry Mutagenesis Molecular Medicine sense organs Protein Binding Subcellular Fractions Biotechnology |
| Zdroj: | Nature Biotechnology. 17:170-175 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/6172 |
| Popis: | PDZ domains are a recently characterized protein-recognition module. In most cases, PDZ domains bind to the C-terminal end of target proteins and are thought thereby to link these target proteins into functional signaling networks. We report the isolation of artificial PDZ domains selected via a mutagenesis screen in vivo, each recognizing a different C-terminal peptide. We demonstrate that the PDZ domains isolated can bind selectively to their target peptides in vitro and in vivo. Two of the target peptides chosen are the C-terminal ends of two cellular transmembrane proteins with which no known PDZ domains have been reported to interact. By targeting these artificial PDZ domains to the nucleus, interacting target peptides were efficiently transported to the same subcellular localization. One of the isolated PDZ domains was tested and shown to be efficiently directed to the plasma membrane when cotransfected with the full-length transmembrane protein in mammalian cells. Thus, artificial PDZ domains can be engineered and used to target intracellular proteins to different subcellular compartments. |
| Databáze: | OpenAIRE |
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