Exploring the Interaction Between the Protein Kinase A Catalytic Subunit and Caveolin-1 Scaffolding Domain with Shotgun Scanning, Oligomer Complementation, Nmr, and Docking.
From: Department of Chemistry, University of California, Irvine, CA 92697, USA.
Protein science : a publication of the Protein Society
- Publish Date: Mar 2006
- ISSN: 0961-8368
- Volume: 15
- Issue: 3
- Pages: 478-86
- Medium: Print
- Language: English
- Citation (JAMA): Levin Aron M, Coroneus John G, Cocco Melanie J, et al. Exploring the Interaction Between the Protein Kinase A Catalytic Subunit and Caveolin-1 Scaffolding Domain with Shotgun Scanning, Oligomer Complementation, Nmr, and Docking.. Protein Sci. Mar 2006;15:478-86
Abstract
The techniques of phage-displayed homolog shotgun scanning, oligomer complementation, NMR secondary structure analysis, and computational docking provide a complementary suite of tools for dissecting protein-protein interactions. Focusing these tools on the interaction between the catalytic sub-unit of protein kinase A (PKAcat) and caveolin-1 scaffolding domain (CSD) reveals the first structural model for the interaction. Homolog shotgun scanning varied each CSD residue as either a wild-type or a homologous amino acid. Wild-type to homolog ratios from 116 different homologous CSD variants identified side-chain functional groups responsible for precise contacts with PKAcat. Structural analysis by NMR assigned an alpha-helical conformation to the central residues 84- 97 of CSD. The extensive mutagenesis data and NMR secondary structure information provided constraints for developing a model for the PKAcat-CSD interaction. Addition of synthetic CSD to phage-displayed CSD resulted in oligomer complementation, or enhanced binding to PKAcat. Together with previous experiments examining the interaction between CSD and endothelial nitric oxide synthase (eNOS), the results suggest a general oligomerization-dependent enhancement of binding between signal transducing enzymes and caveolin-1.
Mesh Headings (Keywords): Binding Sites, Catalytic Domain, Caveolin 1, Computational Biology, Cyclic AMP-Dependent Protein Kinases, Models, Molecular, Mutation, Nuclear Magnetic Resonance, Biomolecular, Peptide Library, Protein Structure, Secondary, Protein Structure, Tertiary, Protein Subunits
Check for Full Text / PubMed Unique Identifier (PMID): 16452625
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