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Analysis of PDZ Domain-Ligand Interactions Using Carboxyl-terminal Phage Display
- Source :
- Journal of Biological Chemistry. 275:21486-21491
- Publication Year :
- 2000
- Publisher :
- Elsevier BV, 2000.
-
Abstract
- PDZ domains mediate protein-protein interactions at specialized subcellular sites, such as epithelial cell tight junctions and neuronal post-synaptic densities. Because most PDZ domains bind extreme carboxyl-terminal sequences, the phage display method has not been amenable to the study of PDZ domain binding specificities. For the first time, we demonstrate the functional display of a peptide library fused to the carboxyl terminus of the M13 major coat protein. We used this library to analyze carboxyl-terminal peptide recognition by two PDZ domains. For each PDZ domain, the library provided specific ligands with sub-micromolar binding affinities. Synthetic peptides and homology modeling were used to dissect and rationalize the binding interactions. Our results establish carboxyl-terminal phage display as a powerful new method for mapping PDZ domain binding specificity.
- Subjects :
- Models, Molecular
Phage display
Protein Conformation
Molecular Sequence Data
PDZ domain
Nerve Tissue Proteins
Biology
Ligands
Biochemistry
Capsid
Protein structure
Peptide Library
Humans
Amino Acid Sequence
Homology modeling
Binding site
Peptide library
Molecular Biology
Peptide sequence
Conserved Sequence
Binding selectivity
Binding Sites
Sequence Homology, Amino Acid
Membrane Proteins
Cell Biology
Molecular biology
Recombinant Proteins
Kinetics
Mutagenesis, Site-Directed
Biophysics
Capsid Proteins
Sequence Alignment
Bacteriophage M13
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 275
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....233f2fc924e0f54d2415629ecf009496