Role of tyrosine 33 residue for the stabilization of the tetrameric structure of human cytidine deaminase
In the present work the effect of a mutation on tyrosine 33 residue (Y33G) of human cytidine deaminase (CDA) was investigated with regard to protein solubility and specific activity. Osmolytes and CDA ligands were used to increase the yield and the specific activity of the protein. The mutant enzyme...
Gespeichert in:
| Veröffentlicht in: | International journal of biological macromolecules 2010-11, Vol.47 (4), p.471-482 |
|---|---|
| Hauptverfasser: | , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 482 |
|---|---|
| container_issue | 4 |
| container_start_page | 471 |
| container_title | International journal of biological macromolecules |
| container_volume | 47 |
| creator | Micozzi, Daniela Pucciarelli, Stefania Carpi, Francesco M. Costanzi, Stefano De Sanctis, Giampiero Polzonetti, Valeria Natalini, Paolo Santarelli, Ivano F. Vita, Alberto Vincenzetti, Silvia |
| description | In the present work the effect of a mutation on tyrosine 33 residue (Y33G) of human cytidine deaminase (CDA) was investigated with regard to protein solubility and specific activity. Osmolytes and CDA ligands were used to increase the yield and the specific activity of the protein. The mutant enzyme was purified and subjected to a kinetic characterization and to stability studies. These investigations reinforced the hypothesis that in human CDA the side chain of Y33 is involved in intersubunit interactions with four glutamate residues (E108) forming a double latch that connects each of the two pairs of monomers of the tetrameric CDA. |
| doi_str_mv | 10.1016/j.ijbiomac.2010.07.001 |
| format | Article |
| fullrecord | <record><control><sourceid>elsevier_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3020594</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0141813010002187</els_id><sourcerecordid>S0141813010002187</sourcerecordid><originalsourceid>FETCH-LOGICAL-c470t-ac4afb0c9f79771f08c61f884b720f71ab94bd0a22af0990f59514cefe7a25273</originalsourceid><addsrcrecordid>eNqFkN1KJDEQhcPiso7uvsLQL9BjJf2T7htRxD8QBNm9Dul0xalhuiNJWhif3oyzM-iVV4FT55yqfIzNOSw48PpstaBVR27QZiEgiSAXAPwHm_FGtjkAFEdsBrzkecMLOGYnIaySWle8-cWOBdSFFKKZMXpya8yczeLGu0AjZkWReQzUT5hZ57O4xCxE3dGa3nQkN36Ykxgxej2gJ5PmfjJx8h9Fy2nQY2Y2kfptXY96oFEH_M1-Wr0O-Of_e8r-3Vz_vbrLHx5v768uH3JTSoi5NqW2HZjWylZKbqExNbdNU3ZSgJVcd23Z9aCF0BbaFmzVVrw0aFFqUQlZnLLzXe_L1A3YGxzTnWv14mnQfqOcJvV1MtJSPbtXVYCAqi1TQb0rMIlI8GgPWQ5qC1-t1B6-2sJXIFWCn4Lzz5sPsT3tZLjYGTD9_5XQq2AIR4M9eTRR9Y6-2_EOrBWcaA</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>Role of tyrosine 33 residue for the stabilization of the tetrameric structure of human cytidine deaminase</title><source>MEDLINE</source><source>ScienceDirect Journals (5 years ago - present)</source><creator>Micozzi, Daniela ; Pucciarelli, Stefania ; Carpi, Francesco M. ; Costanzi, Stefano ; De Sanctis, Giampiero ; Polzonetti, Valeria ; Natalini, Paolo ; Santarelli, Ivano F. ; Vita, Alberto ; Vincenzetti, Silvia</creator><creatorcontrib>Micozzi, Daniela ; Pucciarelli, Stefania ; Carpi, Francesco M. ; Costanzi, Stefano ; De Sanctis, Giampiero ; Polzonetti, Valeria ; Natalini, Paolo ; Santarelli, Ivano F. ; Vita, Alberto ; Vincenzetti, Silvia</creatorcontrib><description>In the present work the effect of a mutation on tyrosine 33 residue (Y33G) of human cytidine deaminase (CDA) was investigated with regard to protein solubility and specific activity. Osmolytes and CDA ligands were used to increase the yield and the specific activity of the protein. The mutant enzyme was purified and subjected to a kinetic characterization and to stability studies. These investigations reinforced the hypothesis that in human CDA the side chain of Y33 is involved in intersubunit interactions with four glutamate residues (E108) forming a double latch that connects each of the two pairs of monomers of the tetrameric CDA.</description><identifier>ISSN: 0141-8130</identifier><identifier>EISSN: 1879-0003</identifier><identifier>DOI: 10.1016/j.ijbiomac.2010.07.001</identifier><identifier>PMID: 20637228</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Blotting, Western ; Catalytic Domain ; Circular Dichroism ; Cytidine deaminase ; Cytidine Deaminase - antagonists & inhibitors ; Cytidine Deaminase - chemistry ; Cytidine Deaminase - isolation & purification ; Cytidine Deaminase - metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability - drug effects ; Humans ; Kinetics ; Mice ; Molecular Chaperones - pharmacology ; Molecular modeling ; Mutant Proteins - antagonists & inhibitors ; Mutant Proteins - isolation & purification ; Mutant Proteins - metabolism ; Protein Structure, Quaternary ; Protein Structure, Secondary ; Protein Unfolding - drug effects ; Site-directed mutagenesis ; Structure-Activity Relationship ; Temperature ; Tyrosine - metabolism</subject><ispartof>International journal of biological macromolecules, 2010-11, Vol.47 (4), p.471-482</ispartof><rights>2010 Elsevier B.V.</rights><rights>Copyright © 2010 Elsevier B.V. All rights reserved.</rights><rights>2010 Elsevier B.V. All rights reserved 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c470t-ac4afb0c9f79771f08c61f884b720f71ab94bd0a22af0990f59514cefe7a25273</citedby><cites>FETCH-LOGICAL-c470t-ac4afb0c9f79771f08c61f884b720f71ab94bd0a22af0990f59514cefe7a25273</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ijbiomac.2010.07.001$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20637228$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Micozzi, Daniela</creatorcontrib><creatorcontrib>Pucciarelli, Stefania</creatorcontrib><creatorcontrib>Carpi, Francesco M.</creatorcontrib><creatorcontrib>Costanzi, Stefano</creatorcontrib><creatorcontrib>De Sanctis, Giampiero</creatorcontrib><creatorcontrib>Polzonetti, Valeria</creatorcontrib><creatorcontrib>Natalini, Paolo</creatorcontrib><creatorcontrib>Santarelli, Ivano F.</creatorcontrib><creatorcontrib>Vita, Alberto</creatorcontrib><creatorcontrib>Vincenzetti, Silvia</creatorcontrib><title>Role of tyrosine 33 residue for the stabilization of the tetrameric structure of human cytidine deaminase</title><title>International journal of biological macromolecules</title><addtitle>Int J Biol Macromol</addtitle><description>In the present work the effect of a mutation on tyrosine 33 residue (Y33G) of human cytidine deaminase (CDA) was investigated with regard to protein solubility and specific activity. Osmolytes and CDA ligands were used to increase the yield and the specific activity of the protein. The mutant enzyme was purified and subjected to a kinetic characterization and to stability studies. These investigations reinforced the hypothesis that in human CDA the side chain of Y33 is involved in intersubunit interactions with four glutamate residues (E108) forming a double latch that connects each of the two pairs of monomers of the tetrameric CDA.</description><subject>Animals</subject><subject>Blotting, Western</subject><subject>Catalytic Domain</subject><subject>Circular Dichroism</subject><subject>Cytidine deaminase</subject><subject>Cytidine Deaminase - antagonists & inhibitors</subject><subject>Cytidine Deaminase - chemistry</subject><subject>Cytidine Deaminase - isolation & purification</subject><subject>Cytidine Deaminase - metabolism</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme Stability - drug effects</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Mice</subject><subject>Molecular Chaperones - pharmacology</subject><subject>Molecular modeling</subject><subject>Mutant Proteins - antagonists & inhibitors</subject><subject>Mutant Proteins - isolation & purification</subject><subject>Mutant Proteins - metabolism</subject><subject>Protein Structure, Quaternary</subject><subject>Protein Structure, Secondary</subject><subject>Protein Unfolding - drug effects</subject><subject>Site-directed mutagenesis</subject><subject>Structure-Activity Relationship</subject><subject>Temperature</subject><subject>Tyrosine - metabolism</subject><issn>0141-8130</issn><issn>1879-0003</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkN1KJDEQhcPiso7uvsLQL9BjJf2T7htRxD8QBNm9Dul0xalhuiNJWhif3oyzM-iVV4FT55yqfIzNOSw48PpstaBVR27QZiEgiSAXAPwHm_FGtjkAFEdsBrzkecMLOGYnIaySWle8-cWOBdSFFKKZMXpya8yczeLGu0AjZkWReQzUT5hZ57O4xCxE3dGa3nQkN36Ykxgxej2gJ5PmfjJx8h9Fy2nQY2Y2kfptXY96oFEH_M1-Wr0O-Of_e8r-3Vz_vbrLHx5v768uH3JTSoi5NqW2HZjWylZKbqExNbdNU3ZSgJVcd23Z9aCF0BbaFmzVVrw0aFFqUQlZnLLzXe_L1A3YGxzTnWv14mnQfqOcJvV1MtJSPbtXVYCAqi1TQb0rMIlI8GgPWQ5qC1-t1B6-2sJXIFWCn4Lzz5sPsT3tZLjYGTD9_5XQq2AIR4M9eTRR9Y6-2_EOrBWcaA</recordid><startdate>20101101</startdate><enddate>20101101</enddate><creator>Micozzi, Daniela</creator><creator>Pucciarelli, Stefania</creator><creator>Carpi, Francesco M.</creator><creator>Costanzi, Stefano</creator><creator>De Sanctis, Giampiero</creator><creator>Polzonetti, Valeria</creator><creator>Natalini, Paolo</creator><creator>Santarelli, Ivano F.</creator><creator>Vita, Alberto</creator><creator>Vincenzetti, Silvia</creator><general>Elsevier B.V</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>5PM</scope></search><sort><creationdate>20101101</creationdate><title>Role of tyrosine 33 residue for the stabilization of the tetrameric structure of human cytidine deaminase</title><author>Micozzi, Daniela ; Pucciarelli, Stefania ; Carpi, Francesco M. ; Costanzi, Stefano ; De Sanctis, Giampiero ; Polzonetti, Valeria ; Natalini, Paolo ; Santarelli, Ivano F. ; Vita, Alberto ; Vincenzetti, Silvia</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c470t-ac4afb0c9f79771f08c61f884b720f71ab94bd0a22af0990f59514cefe7a25273</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animals</topic><topic>Blotting, Western</topic><topic>Catalytic Domain</topic><topic>Circular Dichroism</topic><topic>Cytidine deaminase</topic><topic>Cytidine Deaminase - antagonists & inhibitors</topic><topic>Cytidine Deaminase - chemistry</topic><topic>Cytidine Deaminase - isolation & purification</topic><topic>Cytidine Deaminase - metabolism</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme Stability - drug effects</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Mice</topic><topic>Molecular Chaperones - pharmacology</topic><topic>Molecular modeling</topic><topic>Mutant Proteins - antagonists & inhibitors</topic><topic>Mutant Proteins - isolation & purification</topic><topic>Mutant Proteins - metabolism</topic><topic>Protein Structure, Quaternary</topic><topic>Protein Structure, Secondary</topic><topic>Protein Unfolding - drug effects</topic><topic>Site-directed mutagenesis</topic><topic>Structure-Activity Relationship</topic><topic>Temperature</topic><topic>Tyrosine - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Micozzi, Daniela</creatorcontrib><creatorcontrib>Pucciarelli, Stefania</creatorcontrib><creatorcontrib>Carpi, Francesco M.</creatorcontrib><creatorcontrib>Costanzi, Stefano</creatorcontrib><creatorcontrib>De Sanctis, Giampiero</creatorcontrib><creatorcontrib>Polzonetti, Valeria</creatorcontrib><creatorcontrib>Natalini, Paolo</creatorcontrib><creatorcontrib>Santarelli, Ivano F.</creatorcontrib><creatorcontrib>Vita, Alberto</creatorcontrib><creatorcontrib>Vincenzetti, Silvia</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>International journal of biological macromolecules</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Micozzi, Daniela</au><au>Pucciarelli, Stefania</au><au>Carpi, Francesco M.</au><au>Costanzi, Stefano</au><au>De Sanctis, Giampiero</au><au>Polzonetti, Valeria</au><au>Natalini, Paolo</au><au>Santarelli, Ivano F.</au><au>Vita, Alberto</au><au>Vincenzetti, Silvia</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Role of tyrosine 33 residue for the stabilization of the tetrameric structure of human cytidine deaminase</atitle><jtitle>International journal of biological macromolecules</jtitle><addtitle>Int J Biol Macromol</addtitle><date>2010-11-01</date><risdate>2010</risdate><volume>47</volume><issue>4</issue><spage>471</spage><epage>482</epage><pages>471-482</pages><issn>0141-8130</issn><eissn>1879-0003</eissn><abstract>In the present work the effect of a mutation on tyrosine 33 residue (Y33G) of human cytidine deaminase (CDA) was investigated with regard to protein solubility and specific activity. Osmolytes and CDA ligands were used to increase the yield and the specific activity of the protein. The mutant enzyme was purified and subjected to a kinetic characterization and to stability studies. These investigations reinforced the hypothesis that in human CDA the side chain of Y33 is involved in intersubunit interactions with four glutamate residues (E108) forming a double latch that connects each of the two pairs of monomers of the tetrameric CDA.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>20637228</pmid><doi>10.1016/j.ijbiomac.2010.07.001</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0141-8130 |
| ispartof | International journal of biological macromolecules, 2010-11, Vol.47 (4), p.471-482 |
| issn | 0141-8130 1879-0003 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3020594 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Animals Blotting, Western Catalytic Domain Circular Dichroism Cytidine deaminase Cytidine Deaminase - antagonists & inhibitors Cytidine Deaminase - chemistry Cytidine Deaminase - isolation & purification Cytidine Deaminase - metabolism Electrophoresis, Polyacrylamide Gel Enzyme Stability - drug effects Humans Kinetics Mice Molecular Chaperones - pharmacology Molecular modeling Mutant Proteins - antagonists & inhibitors Mutant Proteins - isolation & purification Mutant Proteins - metabolism Protein Structure, Quaternary Protein Structure, Secondary Protein Unfolding - drug effects Site-directed mutagenesis Structure-Activity Relationship Temperature Tyrosine - metabolism |
| title | Role of tyrosine 33 residue for the stabilization of the tetrameric structure of human cytidine deaminase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-12T07%3A33%3A40IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-elsevier_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Role%20of%20tyrosine%2033%20residue%20for%20the%20stabilization%20of%20the%20tetrameric%20structure%20of%20human%20cytidine%20deaminase&rft.jtitle=International%20journal%20of%20biological%20macromolecules&rft.au=Micozzi,%20Daniela&rft.date=2010-11-01&rft.volume=47&rft.issue=4&rft.spage=471&rft.epage=482&rft.pages=471-482&rft.issn=0141-8130&rft.eissn=1879-0003&rft_id=info:doi/10.1016/j.ijbiomac.2010.07.001&rft_dat=%3Celsevier_pubme%3ES0141813010002187%3C/elsevier_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/20637228&rft_els_id=S0141813010002187&rfr_iscdi=true |