Refolding Parameters for the Allosteric Homodimeric Guanylyl Cyclase Catalytic Core from the Atrial Natriuretic Peptide Receptor
Protein folding continues to be an important biophysical topic in molecular biology. We report the parameters for successfully refolding the guanylyl cyclase core of the ANP receptor, an allosteric homodimeric enzyme. Urea was a better chaotropic solvent than guanidine HCl, and physiological salt co...
Gespeichert in:
| Veröffentlicht in: | Biochemical and biophysical research communications 1996-07, Vol.224 (3), p.765-771 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 771 |
|---|---|
| container_issue | 3 |
| container_start_page | 765 |
| container_title | Biochemical and biophysical research communications |
| container_volume | 224 |
| creator | Thorpe, David S. Niu, Shi Morkin, Eugene |
| description | Protein folding continues to be an important biophysical topic in molecular biology. We report the parameters for successfully refolding the guanylyl cyclase core of the ANP receptor, an allosteric homodimeric enzyme. Urea was a better chaotropic solvent than guanidine HCl, and physiological salt concentrations and pH were needed for optimal recovery of enzymatic activity. Renaturation was more sensitive to alkaline compared to acidic deviations in solvent conditions. The time course of refolding was sigmoidal producing an enzyme with a specific activity of 16,000 pmol cGMP/min/mg using 60 μM concentration of substrate. Additional factors are described in this unusual case of renaturing an allosteric homodimeric enzymein vitro. |
| doi_str_mv | 10.1006/bbrc.1996.1097 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_78190554</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0006291X96910979</els_id><sourcerecordid>78190554</sourcerecordid><originalsourceid>FETCH-LOGICAL-c294t-1e833d964289b492237a49652bcfd0643fd051ca3683f11cbdd6e59dc9b66be03</originalsourceid><addsrcrecordid>eNp1kEtLxDAUhYMoOj627oSs3HVM-sg0Syk6CqIiCu5CmtxqJJ2MSSp05083dQZ3bnJucr57IAehU0rmlBB20bZezSnnLF35YgfNkpAsp6TcRTOSiCzn9PUAHYbwQQilJeP7aL9e0ILmZIa-n6BzVpvVG36UXvYQwQfcOY_jO-BLa11IL0bhG9c7bfrfeTnI1WhHi5tRWRkANzJKO8ZkNc4D7rzrN_vRG2nxvUw6eJiAR1hHowE_gUqT88dor5M2wMlWj9DL9dVzc5PdPSxvm8u7TOW8jBmFuig0Z2Ve87bkeV4sZMlZlbeq04SVRTorqmTB6qKjVLVaM6i4VrxlrAVSHKHzTe7au88BQhS9CQqslStwQxCLmnJSVWUC5xtQeReCh06svemlHwUlYqpcTJWLqXIxVZ4WzrbJQ9uD_sO3HSe_3viQvvdlwIugDKwUaONBRaGd-S_6B7hKkgw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>78190554</pqid></control><display><type>article</type><title>Refolding Parameters for the Allosteric Homodimeric Guanylyl Cyclase Catalytic Core from the Atrial Natriuretic Peptide Receptor</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Thorpe, David S. ; Niu, Shi ; Morkin, Eugene</creator><creatorcontrib>Thorpe, David S. ; Niu, Shi ; Morkin, Eugene</creatorcontrib><description>Protein folding continues to be an important biophysical topic in molecular biology. We report the parameters for successfully refolding the guanylyl cyclase core of the ANP receptor, an allosteric homodimeric enzyme. Urea was a better chaotropic solvent than guanidine HCl, and physiological salt concentrations and pH were needed for optimal recovery of enzymatic activity. Renaturation was more sensitive to alkaline compared to acidic deviations in solvent conditions. The time course of refolding was sigmoidal producing an enzyme with a specific activity of 16,000 pmol cGMP/min/mg using 60 μM concentration of substrate. Additional factors are described in this unusual case of renaturing an allosteric homodimeric enzymein vitro.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1006/bbrc.1996.1097</identifier><identifier>PMID: 8713120</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Allosteric Regulation ; Catalysis ; Cell Line ; Guanylate Cyclase - metabolism ; Hydrogen-Ion Concentration ; Osmolar Concentration ; Phenylmethylsulfonyl Fluoride - pharmacology ; Protein Denaturation ; Protein Folding ; Receptors, Atrial Natriuretic Factor - metabolism ; Salts ; Serine Proteinase Inhibitors - pharmacology ; Solvents ; Substrate Specificity</subject><ispartof>Biochemical and biophysical research communications, 1996-07, Vol.224 (3), p.765-771</ispartof><rights>1996 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/bbrc.1996.1097$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8713120$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Thorpe, David S.</creatorcontrib><creatorcontrib>Niu, Shi</creatorcontrib><creatorcontrib>Morkin, Eugene</creatorcontrib><title>Refolding Parameters for the Allosteric Homodimeric Guanylyl Cyclase Catalytic Core from the Atrial Natriuretic Peptide Receptor</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>Protein folding continues to be an important biophysical topic in molecular biology. We report the parameters for successfully refolding the guanylyl cyclase core of the ANP receptor, an allosteric homodimeric enzyme. Urea was a better chaotropic solvent than guanidine HCl, and physiological salt concentrations and pH were needed for optimal recovery of enzymatic activity. Renaturation was more sensitive to alkaline compared to acidic deviations in solvent conditions. The time course of refolding was sigmoidal producing an enzyme with a specific activity of 16,000 pmol cGMP/min/mg using 60 μM concentration of substrate. Additional factors are described in this unusual case of renaturing an allosteric homodimeric enzymein vitro.</description><subject>Allosteric Regulation</subject><subject>Catalysis</subject><subject>Cell Line</subject><subject>Guanylate Cyclase - metabolism</subject><subject>Hydrogen-Ion Concentration</subject><subject>Osmolar Concentration</subject><subject>Phenylmethylsulfonyl Fluoride - pharmacology</subject><subject>Protein Denaturation</subject><subject>Protein Folding</subject><subject>Receptors, Atrial Natriuretic Factor - metabolism</subject><subject>Salts</subject><subject>Serine Proteinase Inhibitors - pharmacology</subject><subject>Solvents</subject><subject>Substrate Specificity</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kEtLxDAUhYMoOj627oSs3HVM-sg0Syk6CqIiCu5CmtxqJJ2MSSp05083dQZ3bnJucr57IAehU0rmlBB20bZezSnnLF35YgfNkpAsp6TcRTOSiCzn9PUAHYbwQQilJeP7aL9e0ILmZIa-n6BzVpvVG36UXvYQwQfcOY_jO-BLa11IL0bhG9c7bfrfeTnI1WhHi5tRWRkANzJKO8ZkNc4D7rzrN_vRG2nxvUw6eJiAR1hHowE_gUqT88dor5M2wMlWj9DL9dVzc5PdPSxvm8u7TOW8jBmFuig0Z2Ve87bkeV4sZMlZlbeq04SVRTorqmTB6qKjVLVaM6i4VrxlrAVSHKHzTe7au88BQhS9CQqslStwQxCLmnJSVWUC5xtQeReCh06svemlHwUlYqpcTJWLqXIxVZ4WzrbJQ9uD_sO3HSe_3viQvvdlwIugDKwUaONBRaGd-S_6B7hKkgw</recordid><startdate>19960725</startdate><enddate>19960725</enddate><creator>Thorpe, David S.</creator><creator>Niu, Shi</creator><creator>Morkin, Eugene</creator><general>Elsevier Inc</general><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>7X8</scope></search><sort><creationdate>19960725</creationdate><title>Refolding Parameters for the Allosteric Homodimeric Guanylyl Cyclase Catalytic Core from the Atrial Natriuretic Peptide Receptor</title><author>Thorpe, David S. ; Niu, Shi ; Morkin, Eugene</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c294t-1e833d964289b492237a49652bcfd0643fd051ca3683f11cbdd6e59dc9b66be03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Allosteric Regulation</topic><topic>Catalysis</topic><topic>Cell Line</topic><topic>Guanylate Cyclase - metabolism</topic><topic>Hydrogen-Ion Concentration</topic><topic>Osmolar Concentration</topic><topic>Phenylmethylsulfonyl Fluoride - pharmacology</topic><topic>Protein Denaturation</topic><topic>Protein Folding</topic><topic>Receptors, Atrial Natriuretic Factor - metabolism</topic><topic>Salts</topic><topic>Serine Proteinase Inhibitors - pharmacology</topic><topic>Solvents</topic><topic>Substrate Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Thorpe, David S.</creatorcontrib><creatorcontrib>Niu, Shi</creatorcontrib><creatorcontrib>Morkin, Eugene</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Thorpe, David S.</au><au>Niu, Shi</au><au>Morkin, Eugene</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Refolding Parameters for the Allosteric Homodimeric Guanylyl Cyclase Catalytic Core from the Atrial Natriuretic Peptide Receptor</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>1996-07-25</date><risdate>1996</risdate><volume>224</volume><issue>3</issue><spage>765</spage><epage>771</epage><pages>765-771</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>Protein folding continues to be an important biophysical topic in molecular biology. We report the parameters for successfully refolding the guanylyl cyclase core of the ANP receptor, an allosteric homodimeric enzyme. Urea was a better chaotropic solvent than guanidine HCl, and physiological salt concentrations and pH were needed for optimal recovery of enzymatic activity. Renaturation was more sensitive to alkaline compared to acidic deviations in solvent conditions. The time course of refolding was sigmoidal producing an enzyme with a specific activity of 16,000 pmol cGMP/min/mg using 60 μM concentration of substrate. Additional factors are described in this unusual case of renaturing an allosteric homodimeric enzymein vitro.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>8713120</pmid><doi>10.1006/bbrc.1996.1097</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0006-291X |
| ispartof | Biochemical and biophysical research communications, 1996-07, Vol.224 (3), p.765-771 |
| issn | 0006-291X 1090-2104 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_78190554 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Allosteric Regulation Catalysis Cell Line Guanylate Cyclase - metabolism Hydrogen-Ion Concentration Osmolar Concentration Phenylmethylsulfonyl Fluoride - pharmacology Protein Denaturation Protein Folding Receptors, Atrial Natriuretic Factor - metabolism Salts Serine Proteinase Inhibitors - pharmacology Solvents Substrate Specificity |
| title | Refolding Parameters for the Allosteric Homodimeric Guanylyl Cyclase Catalytic Core from the Atrial Natriuretic Peptide Receptor |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-20T17%3A17%3A45IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Refolding%20Parameters%20for%20the%20Allosteric%20Homodimeric%20Guanylyl%20Cyclase%20Catalytic%20Core%20from%20the%20Atrial%20Natriuretic%20Peptide%20Receptor&rft.jtitle=Biochemical%20and%20biophysical%20research%20communications&rft.au=Thorpe,%20David%20S.&rft.date=1996-07-25&rft.volume=224&rft.issue=3&rft.spage=765&rft.epage=771&rft.pages=765-771&rft.issn=0006-291X&rft.eissn=1090-2104&rft_id=info:doi/10.1006/bbrc.1996.1097&rft_dat=%3Cproquest_cross%3E78190554%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=78190554&rft_id=info:pmid/8713120&rft_els_id=S0006291X96910979&rfr_iscdi=true |