Amide H/2H Exchange Reveals Communication Between the cAMP and Catalytic Subunit-binding Sites in the RIα Subunit of Protein Kinase A
| Autor: | Elizabeth A. Komives, Carrie A. Hughes, John M. Jones, Susan S. Taylor, Ganesh S. Anand |
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| Rok vydání: | 2002 |
| Předmět: |
Models
Molecular Stereochemistry Protein subunit Molecular Sequence Data Catalysis chemistry.chemical_compound Structural Biology Catalytic Domain Amide Cyclic AMP Amino Acid Sequence Binding site Protein kinase A Molecular Biology Binding Sites Mutagenesis Amides Cyclic AMP-Dependent Protein Kinases Protein Structure Tertiary Protein Subunits chemistry Biochemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Solvents CAMP binding Signal transduction Hydrogen Protein Binding |
| Zdroj: | Journal of Molecular Biology. 323:377-386 |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/s0022-2836(02)00919-1 |
| Popis: | The changes in backbone hydrogen/deuterium (H/2H) exchange in the regulatory subunit (R(I)alpha(94-244)) of cyclic AMP-dependent protein kinase A (PKA) were probed by MALDI-TOF mass spectrometry. The three naturally occurring states of the regulatory subunit were studied: (1) free R(I)alpha(94-244), which likely represents newly synthesized protein, (2) R(I)alpha(94-244) bound to the catalytic (C) subunit, or holoenzyme, and (3) R(I)alpha(94-244) bound to cAMP. Protection from amide exchange upon C-subunit binding was observed for the helical subdomain, including the A-helix and B-helix, pointing to regions adjacent to those shown to be important by mutagenesis. In addition, C-subunit binding caused changes in observed amide exchange in the distal cAMP-binding pocket. Conversely, cAMP binding caused protection in the cAMP-binding pocket and increased exchange in the helical subdomain. These results suggest that the mutually exclusive binding of either cAMP or C-subunit is controlled by binding at one site transmitting long distance changes to the other site. |
| Databáze: | OpenAIRE |
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