Amide H/2H exchange reveals communication between the cAMP and catalytic subunit-binding sites in the R(I)alpha subunit of protein kinase A
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),...
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| Veröffentlicht in: | Journal of molecular biology 2002-10, Vol.323 (2), p.377 |
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| creator | Anand, Ganesh S Hughes, Carrie A Jones, John M Taylor, Susan S Komives, Elizabeth A |
| description | 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. |
| format | Article |
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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.</description><identifier>ISSN: 0022-2836</identifier><identifier>PMID: 12381327</identifier><language>eng</language><publisher>England</publisher><subject>Amides - chemistry ; Amino Acid Sequence ; Binding Sites ; Catalytic Domain ; Cyclic AMP - chemistry ; Cyclic AMP - metabolism ; Cyclic AMP-Dependent Protein Kinases - chemistry ; Cyclic AMP-Dependent Protein Kinases - metabolism ; Hydrogen - chemistry ; Models, Molecular ; Molecular Sequence Data ; Protein Binding ; Protein Structure, Tertiary ; Protein Subunits - chemistry ; Protein Subunits - metabolism ; Solvents ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><ispartof>Journal of molecular biology, 2002-10, Vol.323 (2), p.377</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12381327$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Anand, Ganesh S</creatorcontrib><creatorcontrib>Hughes, Carrie A</creatorcontrib><creatorcontrib>Jones, John M</creatorcontrib><creatorcontrib>Taylor, Susan S</creatorcontrib><creatorcontrib>Komives, Elizabeth A</creatorcontrib><title>Amide H/2H exchange reveals communication between the cAMP and catalytic subunit-binding sites in the R(I)alpha subunit of protein kinase A</title><title>Journal of molecular biology</title><addtitle>J Mol Biol</addtitle><description>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.</description><subject>Amides - chemistry</subject><subject>Amino Acid Sequence</subject><subject>Binding Sites</subject><subject>Catalytic Domain</subject><subject>Cyclic AMP - chemistry</subject><subject>Cyclic AMP - metabolism</subject><subject>Cyclic AMP-Dependent Protein Kinases - chemistry</subject><subject>Cyclic AMP-Dependent Protein Kinases - metabolism</subject><subject>Hydrogen - chemistry</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Protein Binding</subject><subject>Protein Structure, Tertiary</subject><subject>Protein Subunits - chemistry</subject><subject>Protein Subunits - metabolism</subject><subject>Solvents</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><issn>0022-2836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo1kE1Lw0AYhPeg2Fr9C_Ie9RDczyY9hqK2oCjSe9mPd9vVZhOyW7W_wT9toPY0MPMwA3NGxpRyXvBKTEfkMqUPSqkSsrogI8ZFxQQvx-S3boJDWNzzBeCP3eq4QejxC_UugW2bZh-D1Tm0EQzmb8QIeYtg65c30NHBkOndIQcLaW8GNhcmRBfiBlLImCAc-ffb5Z3edVt9wqD10PVtxgH4DFEnhPqKnPthFq__dUJWjw-r-aJ4fn1azuvnolOyLIxkTFEqtTfcy9nMc82kmErluXIlVdRrLwRa552baVF645kRg0mtohVnYkJujrXd3jTo1l0fGt0f1qdTxB_IKV59</recordid><startdate>20021018</startdate><enddate>20021018</enddate><creator>Anand, Ganesh S</creator><creator>Hughes, Carrie A</creator><creator>Jones, John M</creator><creator>Taylor, Susan S</creator><creator>Komives, Elizabeth A</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope></search><sort><creationdate>20021018</creationdate><title>Amide H/2H exchange reveals communication between the cAMP and catalytic subunit-binding sites in the R(I)alpha subunit of protein kinase A</title><author>Anand, Ganesh S ; Hughes, Carrie A ; Jones, John M ; Taylor, Susan S ; Komives, Elizabeth A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p547-b4115004afb2f499f2a143645f25d7050faf33ecdfdd9a37fbf1b3faf0c508213</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Amides - chemistry</topic><topic>Amino Acid Sequence</topic><topic>Binding Sites</topic><topic>Catalytic Domain</topic><topic>Cyclic AMP - chemistry</topic><topic>Cyclic AMP - metabolism</topic><topic>Cyclic AMP-Dependent Protein Kinases - chemistry</topic><topic>Cyclic AMP-Dependent Protein Kinases - metabolism</topic><topic>Hydrogen - chemistry</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Protein Binding</topic><topic>Protein Structure, Tertiary</topic><topic>Protein Subunits - chemistry</topic><topic>Protein Subunits - metabolism</topic><topic>Solvents</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Anand, Ganesh S</creatorcontrib><creatorcontrib>Hughes, Carrie A</creatorcontrib><creatorcontrib>Jones, John M</creatorcontrib><creatorcontrib>Taylor, Susan S</creatorcontrib><creatorcontrib>Komives, Elizabeth A</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><jtitle>Journal of molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Anand, Ganesh S</au><au>Hughes, Carrie A</au><au>Jones, John M</au><au>Taylor, Susan S</au><au>Komives, Elizabeth A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Amide H/2H exchange reveals communication between the cAMP and catalytic subunit-binding sites in the R(I)alpha subunit of protein kinase A</atitle><jtitle>Journal of molecular biology</jtitle><addtitle>J Mol Biol</addtitle><date>2002-10-18</date><risdate>2002</risdate><volume>323</volume><issue>2</issue><spage>377</spage><pages>377-</pages><issn>0022-2836</issn><abstract>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.</abstract><cop>England</cop><pmid>12381327</pmid></addata></record> |
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| subjects | Amides - chemistry Amino Acid Sequence Binding Sites Catalytic Domain Cyclic AMP - chemistry Cyclic AMP - metabolism Cyclic AMP-Dependent Protein Kinases - chemistry Cyclic AMP-Dependent Protein Kinases - metabolism Hydrogen - chemistry Models, Molecular Molecular Sequence Data Protein Binding Protein Structure, Tertiary Protein Subunits - chemistry Protein Subunits - metabolism Solvents Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| title | Amide H/2H exchange reveals communication between the cAMP and catalytic subunit-binding sites in the R(I)alpha subunit of protein kinase A |
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