Thermodynamics of ribonuclease T1 denaturation
Differential scanning calorimetry has been used to investigate the thermodynamics of denaturation of ribonuclease T1 as a function of pH over the pH range 2-10, and as a function of NaCl and MgCl2 concentration. At pH 7 in 30 mM PIPES buffer, the thermodynamic parameters are as follows: melting temp...
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| Veröffentlicht in: | Biochemistry (Easton) 1992-05, Vol.31 (20), p.4876-4882 |
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| creator | Hu, Cui Qing Sturtevant, Julian M Thomson, James A Erickson, Rick E Pace, C. Nick |
| description | Differential scanning calorimetry has been used to investigate the thermodynamics of denaturation of ribonuclease T1 as a function of pH over the pH range 2-10, and as a function of NaCl and MgCl2 concentration. At pH 7 in 30 mM PIPES buffer, the thermodynamic parameters are as follows: melting temperature, T1/2 = 48.9 +/- 0.1 degrees C; enthalpy change, delta H = 95.5 +/- 0.9 kcal mol-1; heat capacity change, delta Cp = 1.59 kcal mol-1 K-1; free energy change at 25 degrees C, delta G degrees (25 degrees C) = 5.6 kcal mol-1. Both T1/2 = 56.5 degrees C and delta H = 106.1 kcal mol-1 are maximal near pH 5. The conformational stability of ribonuclease T1 is increased by 3.0 kcal/mol in the presence of 0.6 M NaCl or 0.3 M MgCl2. This stabilization results mainly from the preferential binding of cations to the folded conformation of the protein. The estimates of the conformational stability of ribonuclease T1 from differential scanning calorimetry are shown to be in remarkably good agreement with estimates derived from an analysis of urea denaturation curves. |
| doi_str_mv | 10.1021/bi00135a019 |
| format | Article |
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Nick</creator><creatorcontrib>Hu, Cui Qing ; Sturtevant, Julian M ; Thomson, James A ; Erickson, Rick E ; Pace, C. Nick</creatorcontrib><description>Differential scanning calorimetry has been used to investigate the thermodynamics of denaturation of ribonuclease T1 as a function of pH over the pH range 2-10, and as a function of NaCl and MgCl2 concentration. At pH 7 in 30 mM PIPES buffer, the thermodynamic parameters are as follows: melting temperature, T1/2 = 48.9 +/- 0.1 degrees C; enthalpy change, delta H = 95.5 +/- 0.9 kcal mol-1; heat capacity change, delta Cp = 1.59 kcal mol-1 K-1; free energy change at 25 degrees C, delta G degrees (25 degrees C) = 5.6 kcal mol-1. Both T1/2 = 56.5 degrees C and delta H = 106.1 kcal mol-1 are maximal near pH 5. The conformational stability of ribonuclease T1 is increased by 3.0 kcal/mol in the presence of 0.6 M NaCl or 0.3 M MgCl2. This stabilization results mainly from the preferential binding of cations to the folded conformation of the protein. The estimates of the conformational stability of ribonuclease T1 from differential scanning calorimetry are shown to be in remarkably good agreement with estimates derived from an analysis of urea denaturation curves.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi00135a019</identifier><identifier>PMID: 1591247</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Calorimetry, Differential Scanning ; Enzyme Stability ; Hydrogen-Ion Concentration ; Protein Conformation ; Protein Denaturation ; Ribonuclease T1 - chemistry ; Spectrometry, Fluorescence ; Thermodynamics</subject><ispartof>Biochemistry (Easton), 1992-05, Vol.31 (20), p.4876-4882</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a451t-3a5bad3b4dfda468e57fa6bf5aab55abe60f1a955fdc48d00771ff56e1f894d13</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi00135a019$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi00135a019$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1591247$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hu, Cui Qing</creatorcontrib><creatorcontrib>Sturtevant, Julian M</creatorcontrib><creatorcontrib>Thomson, James A</creatorcontrib><creatorcontrib>Erickson, Rick E</creatorcontrib><creatorcontrib>Pace, C. Nick</creatorcontrib><title>Thermodynamics of ribonuclease T1 denaturation</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>Differential scanning calorimetry has been used to investigate the thermodynamics of denaturation of ribonuclease T1 as a function of pH over the pH range 2-10, and as a function of NaCl and MgCl2 concentration. At pH 7 in 30 mM PIPES buffer, the thermodynamic parameters are as follows: melting temperature, T1/2 = 48.9 +/- 0.1 degrees C; enthalpy change, delta H = 95.5 +/- 0.9 kcal mol-1; heat capacity change, delta Cp = 1.59 kcal mol-1 K-1; free energy change at 25 degrees C, delta G degrees (25 degrees C) = 5.6 kcal mol-1. Both T1/2 = 56.5 degrees C and delta H = 106.1 kcal mol-1 are maximal near pH 5. The conformational stability of ribonuclease T1 is increased by 3.0 kcal/mol in the presence of 0.6 M NaCl or 0.3 M MgCl2. This stabilization results mainly from the preferential binding of cations to the folded conformation of the protein. The estimates of the conformational stability of ribonuclease T1 from differential scanning calorimetry are shown to be in remarkably good agreement with estimates derived from an analysis of urea denaturation curves.</description><subject>Calorimetry, Differential Scanning</subject><subject>Enzyme Stability</subject><subject>Hydrogen-Ion Concentration</subject><subject>Protein Conformation</subject><subject>Protein Denaturation</subject><subject>Ribonuclease T1 - chemistry</subject><subject>Spectrometry, Fluorescence</subject><subject>Thermodynamics</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1Lw0AQhhdRaq2ePAs96UFSd5L9SI5a1ApFRSN4WybZXUzNh-4mYP-9kRT1IHgahvfhHeYh5BDoDGgIZ1lBKUQcKSRbZAw8pAFLEr5NxpRSEYSJoLtkz_tVvzIq2YiMgCcQMjkms_TFuKrR6xqrIvfTxk5dkTV1l5cGvZmmMNWmxrZz2BZNvU92LJbeHGzmhDxdXabzRbC8u76Zny8DZBzaIEKeoY4ypq1GJmLDpUWRWY6YcY6ZEdQCJpxbnbNYUyolWMuFARsnTEM0IcdD75tr3jvjW1UVPjdlibVpOq9k_xSLJfsXBCEY9GQPng5g7hrvnbHqzRUVurUCqr40ql8ae_poU9tlldE_7OCtz4MhL3xrPr5jdK9KyEhyld4_qmf5IG8XS64uev5k4DH3atV0ru7t_Xn5EyE4iFg</recordid><startdate>19920501</startdate><enddate>19920501</enddate><creator>Hu, Cui Qing</creator><creator>Sturtevant, Julian M</creator><creator>Thomson, James A</creator><creator>Erickson, Rick E</creator><creator>Pace, C. Nick</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>19920501</creationdate><title>Thermodynamics of ribonuclease T1 denaturation</title><author>Hu, Cui Qing ; Sturtevant, Julian M ; Thomson, James A ; Erickson, Rick E ; Pace, C. Nick</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a451t-3a5bad3b4dfda468e57fa6bf5aab55abe60f1a955fdc48d00771ff56e1f894d13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Calorimetry, Differential Scanning</topic><topic>Enzyme Stability</topic><topic>Hydrogen-Ion Concentration</topic><topic>Protein Conformation</topic><topic>Protein Denaturation</topic><topic>Ribonuclease T1 - chemistry</topic><topic>Spectrometry, Fluorescence</topic><topic>Thermodynamics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hu, Cui Qing</creatorcontrib><creatorcontrib>Sturtevant, Julian M</creatorcontrib><creatorcontrib>Thomson, James A</creatorcontrib><creatorcontrib>Erickson, Rick E</creatorcontrib><creatorcontrib>Pace, C. Nick</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hu, Cui Qing</au><au>Sturtevant, Julian M</au><au>Thomson, James A</au><au>Erickson, Rick E</au><au>Pace, C. Nick</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Thermodynamics of ribonuclease T1 denaturation</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>1992-05-01</date><risdate>1992</risdate><volume>31</volume><issue>20</issue><spage>4876</spage><epage>4882</epage><pages>4876-4882</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>Differential scanning calorimetry has been used to investigate the thermodynamics of denaturation of ribonuclease T1 as a function of pH over the pH range 2-10, and as a function of NaCl and MgCl2 concentration. At pH 7 in 30 mM PIPES buffer, the thermodynamic parameters are as follows: melting temperature, T1/2 = 48.9 +/- 0.1 degrees C; enthalpy change, delta H = 95.5 +/- 0.9 kcal mol-1; heat capacity change, delta Cp = 1.59 kcal mol-1 K-1; free energy change at 25 degrees C, delta G degrees (25 degrees C) = 5.6 kcal mol-1. Both T1/2 = 56.5 degrees C and delta H = 106.1 kcal mol-1 are maximal near pH 5. The conformational stability of ribonuclease T1 is increased by 3.0 kcal/mol in the presence of 0.6 M NaCl or 0.3 M MgCl2. This stabilization results mainly from the preferential binding of cations to the folded conformation of the protein. The estimates of the conformational stability of ribonuclease T1 from differential scanning calorimetry are shown to be in remarkably good agreement with estimates derived from an analysis of urea denaturation curves.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>1591247</pmid><doi>10.1021/bi00135a019</doi><tpages>7</tpages></addata></record> |
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| source | MEDLINE; American Chemical Society Journals |
| subjects | Calorimetry, Differential Scanning Enzyme Stability Hydrogen-Ion Concentration Protein Conformation Protein Denaturation Ribonuclease T1 - chemistry Spectrometry, Fluorescence Thermodynamics |
| title | Thermodynamics of ribonuclease T1 denaturation |
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