Molten Globule-Like State of Bovine Carbonic Anhydrase in the Presence of Acetonitrile
We have evaluated the effects of acetonitrile on the structure and function of bovine carbonic anhydrase II. The potential structural and functional changes in carbonic anhydrase in the presence of different acetonitrile/buffer ratios (0%, 17.5% and 47.5% v/v) were determined using a variety of meth...
Gespeichert in:
| Veröffentlicht in: | Journal of biochemistry (Tokyo) 2006-06, Vol.139 (6), p.1025-1033 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1033 |
|---|---|
| container_issue | 6 |
| container_start_page | 1025 |
| container_title | Journal of biochemistry (Tokyo) |
| container_volume | 139 |
| creator | Safarian, Shahrokh Saffarzadeh, Mona Zargar, Sayyed Jalal Moosavi-Movahedi, Ali Akbar |
| description | We have evaluated the effects of acetonitrile on the structure and function of bovine carbonic anhydrase II. The potential structural and functional changes in carbonic anhydrase in the presence of different acetonitrile/buffer ratios (0%, 17.5% and 47.5% v/v) were determined using a variety of methods. These included simple spectrophotometric methods to record enzyme velocity, fluorescence measurements and calculation of accessible surface area (ASA) to identify possible alterations in tertiary structure of the protein, CD measurements to search for secondary structure conversions, and thermal scanning to determine structural stability of the protein in different media. The Far-UV CD studies indicated that carbonic anhydrase, for the most part, retains its secondary structure in the presence of acetonitrile. Fluorescence measurements using iodide ion and ANS along with ASA calculations revealed that in the presence of acetonitrile some degree of conformational change occurs in the carbonic anhydrase structure. In addition to the hydrophobic pockets, two additional tryptophanyl residues become exposed to the solvent, thereby increasing the surface hydrophobicity of the protein. These alterations dramatically reduce the catalytic activity, thermal stability, and aggregation velocity of the enzyme. Thus, our results support a molten globule-like structure of carbonic anhydrase in the presence of acetonitrile. |
| doi_str_mv | 10.1093/jb/mvj115 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_68108853</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>17261183</sourcerecordid><originalsourceid>FETCH-LOGICAL-c425t-e77e30449c26514cff211705836fd8701d23430bf141a0382ee86c7ee379efac3</originalsourceid><addsrcrecordid>eNqF0MFuEzEQBmALgWhaOPAC4BNSD0s99q7tHNMIWiARSKUQcbG8zpg63ayL7VT07dmwERw5jUbzaTTzE_IC2BtgU3G2ac-29xuA5hGZgGpkxWUDj8mEMQ7VlNerI3Kc82bfciGekiOQSmvWiAn5uoxdwZ5edLHddVgtwi3Sq2IL0ujpebwPPdK5TW3sg6Oz_uZhnWxGGnpabpB-Tpixd3_wzGEZVEmhw2fkibddxueHekKu3739Mr-sFp8u3s9ni8rVvCkVKoWC1fXU7S-unfccQLFGC-nXWjFYc1EL1nqowTKhOaKWTiEKNUVvnTghr8e9dyn-3GEuZhuyw66zPcZdNlID07oR_4WguATQe3g6Qpdizgm9uUtha9ODAWb2aZtNa8a0B_vysHTXbnH9Tx7iHUA1gpAL_vo7t-nWSCVUYy5X3438tlLLD_VHsxz8q9F7G439kUI211ecgWDDG0IIKX4DpImSVA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17261183</pqid></control><display><type>article</type><title>Molten Globule-Like State of Bovine Carbonic Anhydrase in the Presence of Acetonitrile</title><source>MEDLINE</source><source>Oxford University Press Journals All Titles (1996-Current)</source><creator>Safarian, Shahrokh ; Saffarzadeh, Mona ; Zargar, Sayyed Jalal ; Moosavi-Movahedi, Ali Akbar</creator><creatorcontrib>Safarian, Shahrokh ; Saffarzadeh, Mona ; Zargar, Sayyed Jalal ; Moosavi-Movahedi, Ali Akbar</creatorcontrib><description>We have evaluated the effects of acetonitrile on the structure and function of bovine carbonic anhydrase II. The potential structural and functional changes in carbonic anhydrase in the presence of different acetonitrile/buffer ratios (0%, 17.5% and 47.5% v/v) were determined using a variety of methods. These included simple spectrophotometric methods to record enzyme velocity, fluorescence measurements and calculation of accessible surface area (ASA) to identify possible alterations in tertiary structure of the protein, CD measurements to search for secondary structure conversions, and thermal scanning to determine structural stability of the protein in different media. The Far-UV CD studies indicated that carbonic anhydrase, for the most part, retains its secondary structure in the presence of acetonitrile. Fluorescence measurements using iodide ion and ANS along with ASA calculations revealed that in the presence of acetonitrile some degree of conformational change occurs in the carbonic anhydrase structure. In addition to the hydrophobic pockets, two additional tryptophanyl residues become exposed to the solvent, thereby increasing the surface hydrophobicity of the protein. These alterations dramatically reduce the catalytic activity, thermal stability, and aggregation velocity of the enzyme. Thus, our results support a molten globule-like structure of carbonic anhydrase in the presence of acetonitrile.</description><identifier>ISSN: 0021-924X</identifier><identifier>EISSN: 1756-2651</identifier><identifier>DOI: 10.1093/jb/mvj115</identifier><identifier>PMID: 16788053</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>a.u ; accessible surface area ; acetonitrile ; Acetonitriles - pharmacology ; anilinonaphtalene sulfonic acid ; Animals ; ANS ; arbitrary unit ; ASA ; Å2 ; carbonic anhydrase ; Carbonic Anhydrase II - chemistry ; Carbonic Anhydrase II - metabolism ; Carbonic Anhydrases - chemistry ; Carbonic Anhydrases - metabolism ; Catalysis - drug effects ; Cattle ; circular dichroism ; Circular Dichroism - methods ; Dose-Response Relationship, Drug ; Enzyme Stability - drug effects ; extinction coefficient ; His ; histidine ; Kinetics ; mAb/min ; milliabsorbance per minute ; molten globule ; organic solvent ; p-nitrophenylacetate ; p-NPA ; potassium iodide ; Protein Conformation - drug effects ; Protein Folding ; Spectrometry, Fluorescence - methods ; Structure-Activity Relationship ; surface unit equal to one square of Angstrom ; tetrahydrofuran ; thermal stability ; THF ; Trp ; tryptophan ; Tryptophan - chemistry ; v/v ; volume per volume</subject><ispartof>Journal of biochemistry (Tokyo), 2006-06, Vol.139 (6), p.1025-1033</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c425t-e77e30449c26514cff211705836fd8701d23430bf141a0382ee86c7ee379efac3</citedby><cites>FETCH-LOGICAL-c425t-e77e30449c26514cff211705836fd8701d23430bf141a0382ee86c7ee379efac3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16788053$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Safarian, Shahrokh</creatorcontrib><creatorcontrib>Saffarzadeh, Mona</creatorcontrib><creatorcontrib>Zargar, Sayyed Jalal</creatorcontrib><creatorcontrib>Moosavi-Movahedi, Ali Akbar</creatorcontrib><title>Molten Globule-Like State of Bovine Carbonic Anhydrase in the Presence of Acetonitrile</title><title>Journal of biochemistry (Tokyo)</title><addtitle>J Biochem</addtitle><description>We have evaluated the effects of acetonitrile on the structure and function of bovine carbonic anhydrase II. The potential structural and functional changes in carbonic anhydrase in the presence of different acetonitrile/buffer ratios (0%, 17.5% and 47.5% v/v) were determined using a variety of methods. These included simple spectrophotometric methods to record enzyme velocity, fluorescence measurements and calculation of accessible surface area (ASA) to identify possible alterations in tertiary structure of the protein, CD measurements to search for secondary structure conversions, and thermal scanning to determine structural stability of the protein in different media. The Far-UV CD studies indicated that carbonic anhydrase, for the most part, retains its secondary structure in the presence of acetonitrile. Fluorescence measurements using iodide ion and ANS along with ASA calculations revealed that in the presence of acetonitrile some degree of conformational change occurs in the carbonic anhydrase structure. In addition to the hydrophobic pockets, two additional tryptophanyl residues become exposed to the solvent, thereby increasing the surface hydrophobicity of the protein. These alterations dramatically reduce the catalytic activity, thermal stability, and aggregation velocity of the enzyme. Thus, our results support a molten globule-like structure of carbonic anhydrase in the presence of acetonitrile.</description><subject>a.u</subject><subject>accessible surface area</subject><subject>acetonitrile</subject><subject>Acetonitriles - pharmacology</subject><subject>anilinonaphtalene sulfonic acid</subject><subject>Animals</subject><subject>ANS</subject><subject>arbitrary unit</subject><subject>ASA</subject><subject>Å2</subject><subject>carbonic anhydrase</subject><subject>Carbonic Anhydrase II - chemistry</subject><subject>Carbonic Anhydrase II - metabolism</subject><subject>Carbonic Anhydrases - chemistry</subject><subject>Carbonic Anhydrases - metabolism</subject><subject>Catalysis - drug effects</subject><subject>Cattle</subject><subject>circular dichroism</subject><subject>Circular Dichroism - methods</subject><subject>Dose-Response Relationship, Drug</subject><subject>Enzyme Stability - drug effects</subject><subject>extinction coefficient</subject><subject>His</subject><subject>histidine</subject><subject>Kinetics</subject><subject>mAb/min</subject><subject>milliabsorbance per minute</subject><subject>molten globule</subject><subject>organic solvent</subject><subject>p-nitrophenylacetate</subject><subject>p-NPA</subject><subject>potassium iodide</subject><subject>Protein Conformation - drug effects</subject><subject>Protein Folding</subject><subject>Spectrometry, Fluorescence - methods</subject><subject>Structure-Activity Relationship</subject><subject>surface unit equal to one square of Angstrom</subject><subject>tetrahydrofuran</subject><subject>thermal stability</subject><subject>THF</subject><subject>Trp</subject><subject>tryptophan</subject><subject>Tryptophan - chemistry</subject><subject>v/v</subject><subject>volume per volume</subject><issn>0021-924X</issn><issn>1756-2651</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0MFuEzEQBmALgWhaOPAC4BNSD0s99q7tHNMIWiARSKUQcbG8zpg63ayL7VT07dmwERw5jUbzaTTzE_IC2BtgU3G2ac-29xuA5hGZgGpkxWUDj8mEMQ7VlNerI3Kc82bfciGekiOQSmvWiAn5uoxdwZ5edLHddVgtwi3Sq2IL0ujpebwPPdK5TW3sg6Oz_uZhnWxGGnpabpB-Tpixd3_wzGEZVEmhw2fkibddxueHekKu3739Mr-sFp8u3s9ni8rVvCkVKoWC1fXU7S-unfccQLFGC-nXWjFYc1EL1nqowTKhOaKWTiEKNUVvnTghr8e9dyn-3GEuZhuyw66zPcZdNlID07oR_4WguATQe3g6Qpdizgm9uUtha9ODAWb2aZtNa8a0B_vysHTXbnH9Tx7iHUA1gpAL_vo7t-nWSCVUYy5X3438tlLLD_VHsxz8q9F7G439kUI211ecgWDDG0IIKX4DpImSVA</recordid><startdate>20060601</startdate><enddate>20060601</enddate><creator>Safarian, Shahrokh</creator><creator>Saffarzadeh, Mona</creator><creator>Zargar, Sayyed Jalal</creator><creator>Moosavi-Movahedi, Ali Akbar</creator><general>Oxford University Press</general><scope>FBQ</scope><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>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20060601</creationdate><title>Molten Globule-Like State of Bovine Carbonic Anhydrase in the Presence of Acetonitrile</title><author>Safarian, Shahrokh ; Saffarzadeh, Mona ; Zargar, Sayyed Jalal ; Moosavi-Movahedi, Ali Akbar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c425t-e77e30449c26514cff211705836fd8701d23430bf141a0382ee86c7ee379efac3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>a.u</topic><topic>accessible surface area</topic><topic>acetonitrile</topic><topic>Acetonitriles - pharmacology</topic><topic>anilinonaphtalene sulfonic acid</topic><topic>Animals</topic><topic>ANS</topic><topic>arbitrary unit</topic><topic>ASA</topic><topic>Å2</topic><topic>carbonic anhydrase</topic><topic>Carbonic Anhydrase II - chemistry</topic><topic>Carbonic Anhydrase II - metabolism</topic><topic>Carbonic Anhydrases - chemistry</topic><topic>Carbonic Anhydrases - metabolism</topic><topic>Catalysis - drug effects</topic><topic>Cattle</topic><topic>circular dichroism</topic><topic>Circular Dichroism - methods</topic><topic>Dose-Response Relationship, Drug</topic><topic>Enzyme Stability - drug effects</topic><topic>extinction coefficient</topic><topic>His</topic><topic>histidine</topic><topic>Kinetics</topic><topic>mAb/min</topic><topic>milliabsorbance per minute</topic><topic>molten globule</topic><topic>organic solvent</topic><topic>p-nitrophenylacetate</topic><topic>p-NPA</topic><topic>potassium iodide</topic><topic>Protein Conformation - drug effects</topic><topic>Protein Folding</topic><topic>Spectrometry, Fluorescence - methods</topic><topic>Structure-Activity Relationship</topic><topic>surface unit equal to one square of Angstrom</topic><topic>tetrahydrofuran</topic><topic>thermal stability</topic><topic>THF</topic><topic>Trp</topic><topic>tryptophan</topic><topic>Tryptophan - chemistry</topic><topic>v/v</topic><topic>volume per volume</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Safarian, Shahrokh</creatorcontrib><creatorcontrib>Saffarzadeh, Mona</creatorcontrib><creatorcontrib>Zargar, Sayyed Jalal</creatorcontrib><creatorcontrib>Moosavi-Movahedi, Ali Akbar</creatorcontrib><collection>AGRIS</collection><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>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biochemistry (Tokyo)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Safarian, Shahrokh</au><au>Saffarzadeh, Mona</au><au>Zargar, Sayyed Jalal</au><au>Moosavi-Movahedi, Ali Akbar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molten Globule-Like State of Bovine Carbonic Anhydrase in the Presence of Acetonitrile</atitle><jtitle>Journal of biochemistry (Tokyo)</jtitle><addtitle>J Biochem</addtitle><date>2006-06-01</date><risdate>2006</risdate><volume>139</volume><issue>6</issue><spage>1025</spage><epage>1033</epage><pages>1025-1033</pages><issn>0021-924X</issn><eissn>1756-2651</eissn><abstract>We have evaluated the effects of acetonitrile on the structure and function of bovine carbonic anhydrase II. The potential structural and functional changes in carbonic anhydrase in the presence of different acetonitrile/buffer ratios (0%, 17.5% and 47.5% v/v) were determined using a variety of methods. These included simple spectrophotometric methods to record enzyme velocity, fluorescence measurements and calculation of accessible surface area (ASA) to identify possible alterations in tertiary structure of the protein, CD measurements to search for secondary structure conversions, and thermal scanning to determine structural stability of the protein in different media. The Far-UV CD studies indicated that carbonic anhydrase, for the most part, retains its secondary structure in the presence of acetonitrile. Fluorescence measurements using iodide ion and ANS along with ASA calculations revealed that in the presence of acetonitrile some degree of conformational change occurs in the carbonic anhydrase structure. In addition to the hydrophobic pockets, two additional tryptophanyl residues become exposed to the solvent, thereby increasing the surface hydrophobicity of the protein. These alterations dramatically reduce the catalytic activity, thermal stability, and aggregation velocity of the enzyme. Thus, our results support a molten globule-like structure of carbonic anhydrase in the presence of acetonitrile.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>16788053</pmid><doi>10.1093/jb/mvj115</doi><tpages>9</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-924X |
| ispartof | Journal of biochemistry (Tokyo), 2006-06, Vol.139 (6), p.1025-1033 |
| issn | 0021-924X 1756-2651 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_68108853 |
| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current) |
| subjects | a.u accessible surface area acetonitrile Acetonitriles - pharmacology anilinonaphtalene sulfonic acid Animals ANS arbitrary unit ASA Å2 carbonic anhydrase Carbonic Anhydrase II - chemistry Carbonic Anhydrase II - metabolism Carbonic Anhydrases - chemistry Carbonic Anhydrases - metabolism Catalysis - drug effects Cattle circular dichroism Circular Dichroism - methods Dose-Response Relationship, Drug Enzyme Stability - drug effects extinction coefficient His histidine Kinetics mAb/min milliabsorbance per minute molten globule organic solvent p-nitrophenylacetate p-NPA potassium iodide Protein Conformation - drug effects Protein Folding Spectrometry, Fluorescence - methods Structure-Activity Relationship surface unit equal to one square of Angstrom tetrahydrofuran thermal stability THF Trp tryptophan Tryptophan - chemistry v/v volume per volume |
| title | Molten Globule-Like State of Bovine Carbonic Anhydrase in the Presence of Acetonitrile |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-19T08%3A50%3A51IST&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=Molten%20Globule-Like%20State%20of%20Bovine%20Carbonic%20Anhydrase%20in%20the%20Presence%20of%20Acetonitrile&rft.jtitle=Journal%20of%20biochemistry%20(Tokyo)&rft.au=Safarian,%20Shahrokh&rft.date=2006-06-01&rft.volume=139&rft.issue=6&rft.spage=1025&rft.epage=1033&rft.pages=1025-1033&rft.issn=0021-924X&rft.eissn=1756-2651&rft_id=info:doi/10.1093/jb/mvj115&rft_dat=%3Cproquest_cross%3E17261183%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=17261183&rft_id=info:pmid/16788053&rfr_iscdi=true |