Involvement of residues 296–299 in the enzymatic activity of tissue-type plasminogen activator
The tetra-alanine substitution variant KHRR 296–299 AAAA of tissue-type plasminogen activator (t-PA) was previously shown to have enhanced fibrin specificity and enhanced activity in the presence of fibrin compared with the wild-type form of the molecule. The structural requirements for these altera...
Gespeichert in:
| Veröffentlicht in: | Protein engineering 1992-04, Vol.5 (3), p.259-266 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 266 |
|---|---|
| container_issue | 3 |
| container_start_page | 259 |
| container_title | Protein engineering |
| container_volume | 5 |
| creator | Paoni, Nicholas F. Refino, Canio J. Brady, Kevin Pena, Luis C. Nguyen, Hung V. Kerr, Elizabeth M. Johnson, Adriana C. Wurm, Florian M. van Reis, Robert Botstein, David Bennett, William F. |
| description | The tetra-alanine substitution variant KHRR 296–299 AAAA of tissue-type plasminogen activator (t-PA) was previously shown to have enhanced fibrin specificity and enhanced activity in the presence of fibrin compared with the wild-type form of the molecule. The structural requirements for these alterations in enzymatic activity were investigated by constructing several amino acid substitution variants at each of the positions from 296 to 299 and evaluating their activities under a variety of conditions. Effects on plasminogen activator activity were common among the point mutants at positions 296–299; nearly all had a phenotype similar to the KHRR 296–299 AAAA variant. The greatest effects on enzymatic function were found with multiple substitution variants, but some single charge reversals and proline substitutions had substantial effects. The enhanced fibrin specificity of KHRR 296–299 AAAA t-PA results in less fibrinogenolysis than seen with wild-type t-PA. Approximately four times greater concentration of KHRR 296–299 AAAA compared with wild-type t-PA was required to consume 50% of the fibrinogen in human plasma. |
| doi_str_mv | 10.1093/protein/5.3.259 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_16501377</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>16501377</sourcerecordid><originalsourceid>FETCH-LOGICAL-c390t-95db9541038030f0561fb3535c1592bfdb2318b0c1353590f59c8f384cc9bc013</originalsourceid><addsrcrecordid>eNpFkMFOAjEQhhujQUTPnkz2YLwttNvtLj0aFCFiDIka46V2S6vV3S62hYgn38E39EksWYKnTvp_M5n5ADhGsIsgxb25rb3Upke6uJsQugPaKE9RDBFOd7d1ku2DA-feIEyyHKEWaKEUUpLmbfA8Nsu6XMpKGh_VKrLS6dlCuiih2e_3T0JppE3kX2Ukzdeq4l6LiAuvl9qv1rzXzi1k7FdzGc1L7ipt6hdpGob72h6CPcVLJ482bwfcDy_vBqN4cns1HpxPYoEp9DElsyJshCDuQwwVJBlSBSaYCERoUqhZkWDUL6BA608KFaGir3A_FYIWIpzbAWfN3GDkIxzgWaWdkGXJjawXjqGMBCrPA9hrQGFr56xUbG51xe2KIcjWTtnGKSMMs-A0dJxsRi-KSs7--UZiyE83OXeCl8pyI7TbYikmYWUYsLjBtPPycxtz-86yHOeEjR6fGL6ZXDxMh9dsiv8AH6-QsA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16501377</pqid></control><display><type>article</type><title>Involvement of residues 296–299 in the enzymatic activity of tissue-type plasminogen activator</title><source>MEDLINE</source><source>Alma/SFX Local Collection</source><source>Oxford University Press Journals Digital Archive Legacy</source><creator>Paoni, Nicholas F. ; Refino, Canio J. ; Brady, Kevin ; Pena, Luis C. ; Nguyen, Hung V. ; Kerr, Elizabeth M. ; Johnson, Adriana C. ; Wurm, Florian M. ; van Reis, Robert ; Botstein, David ; Bennett, William F.</creator><creatorcontrib>Paoni, Nicholas F. ; Refino, Canio J. ; Brady, Kevin ; Pena, Luis C. ; Nguyen, Hung V. ; Kerr, Elizabeth M. ; Johnson, Adriana C. ; Wurm, Florian M. ; van Reis, Robert ; Botstein, David ; Bennett, William F.</creatorcontrib><description>The tetra-alanine substitution variant KHRR 296–299 AAAA of tissue-type plasminogen activator (t-PA) was previously shown to have enhanced fibrin specificity and enhanced activity in the presence of fibrin compared with the wild-type form of the molecule. The structural requirements for these alterations in enzymatic activity were investigated by constructing several amino acid substitution variants at each of the positions from 296 to 299 and evaluating their activities under a variety of conditions. Effects on plasminogen activator activity were common among the point mutants at positions 296–299; nearly all had a phenotype similar to the KHRR 296–299 AAAA variant. The greatest effects on enzymatic function were found with multiple substitution variants, but some single charge reversals and proline substitutions had substantial effects. The enhanced fibrin specificity of KHRR 296–299 AAAA t-PA results in less fibrinogenolysis than seen with wild-type t-PA. Approximately four times greater concentration of KHRR 296–299 AAAA compared with wild-type t-PA was required to consume 50% of the fibrinogen in human plasma.</description><identifier>ISSN: 1741-0126</identifier><identifier>ISSN: 0269-2139</identifier><identifier>EISSN: 1741-0134</identifier><identifier>EISSN: 1460-213X</identifier><identifier>DOI: 10.1093/protein/5.3.259</identifier><identifier>PMID: 1409547</identifier><identifier>CODEN: PRENE9</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>Alanine - chemistry ; Amino Acid Sequence ; Analytical, structural and metabolic biochemistry ; Biological and medical sciences ; Enzymes and enzyme inhibitors ; fibrin ; Fibrin - metabolism ; Fibrinogen - metabolism ; Fibrinolysis ; Fundamental and applied biological sciences. Psychology ; man ; Models, Molecular ; Molecular Sequence Data ; mutagenesis ; Mutagenesis, Site-Directed ; Oligopeptides - metabolism ; Plasma ; Plasminogen - metabolism ; plasminogen activator ; Protein Engineering ; site-directed mutagenesis ; specificity ; Tissue Plasminogen Activator - chemistry ; Tissue Plasminogen Activator - metabolism ; tissue-type plasminogen activator</subject><ispartof>Protein engineering, 1992-04, Vol.5 (3), p.259-266</ispartof><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c390t-95db9541038030f0561fb3535c1592bfdb2318b0c1353590f59c8f384cc9bc013</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4355350$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1409547$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Paoni, Nicholas F.</creatorcontrib><creatorcontrib>Refino, Canio J.</creatorcontrib><creatorcontrib>Brady, Kevin</creatorcontrib><creatorcontrib>Pena, Luis C.</creatorcontrib><creatorcontrib>Nguyen, Hung V.</creatorcontrib><creatorcontrib>Kerr, Elizabeth M.</creatorcontrib><creatorcontrib>Johnson, Adriana C.</creatorcontrib><creatorcontrib>Wurm, Florian M.</creatorcontrib><creatorcontrib>van Reis, Robert</creatorcontrib><creatorcontrib>Botstein, David</creatorcontrib><creatorcontrib>Bennett, William F.</creatorcontrib><title>Involvement of residues 296–299 in the enzymatic activity of tissue-type plasminogen activator</title><title>Protein engineering</title><addtitle>Protein Eng</addtitle><description>The tetra-alanine substitution variant KHRR 296–299 AAAA of tissue-type plasminogen activator (t-PA) was previously shown to have enhanced fibrin specificity and enhanced activity in the presence of fibrin compared with the wild-type form of the molecule. The structural requirements for these alterations in enzymatic activity were investigated by constructing several amino acid substitution variants at each of the positions from 296 to 299 and evaluating their activities under a variety of conditions. Effects on plasminogen activator activity were common among the point mutants at positions 296–299; nearly all had a phenotype similar to the KHRR 296–299 AAAA variant. The greatest effects on enzymatic function were found with multiple substitution variants, but some single charge reversals and proline substitutions had substantial effects. The enhanced fibrin specificity of KHRR 296–299 AAAA t-PA results in less fibrinogenolysis than seen with wild-type t-PA. Approximately four times greater concentration of KHRR 296–299 AAAA compared with wild-type t-PA was required to consume 50% of the fibrinogen in human plasma.</description><subject>Alanine - chemistry</subject><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Enzymes and enzyme inhibitors</subject><subject>fibrin</subject><subject>Fibrin - metabolism</subject><subject>Fibrinogen - metabolism</subject><subject>Fibrinolysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>man</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>mutagenesis</subject><subject>Mutagenesis, Site-Directed</subject><subject>Oligopeptides - metabolism</subject><subject>Plasma</subject><subject>Plasminogen - metabolism</subject><subject>plasminogen activator</subject><subject>Protein Engineering</subject><subject>site-directed mutagenesis</subject><subject>specificity</subject><subject>Tissue Plasminogen Activator - chemistry</subject><subject>Tissue Plasminogen Activator - metabolism</subject><subject>tissue-type plasminogen activator</subject><issn>1741-0126</issn><issn>0269-2139</issn><issn>1741-0134</issn><issn>1460-213X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkMFOAjEQhhujQUTPnkz2YLwttNvtLj0aFCFiDIka46V2S6vV3S62hYgn38E39EksWYKnTvp_M5n5ADhGsIsgxb25rb3Upke6uJsQugPaKE9RDBFOd7d1ku2DA-feIEyyHKEWaKEUUpLmbfA8Nsu6XMpKGh_VKrLS6dlCuiih2e_3T0JppE3kX2Ukzdeq4l6LiAuvl9qv1rzXzi1k7FdzGc1L7ipt6hdpGob72h6CPcVLJ482bwfcDy_vBqN4cns1HpxPYoEp9DElsyJshCDuQwwVJBlSBSaYCERoUqhZkWDUL6BA608KFaGir3A_FYIWIpzbAWfN3GDkIxzgWaWdkGXJjawXjqGMBCrPA9hrQGFr56xUbG51xe2KIcjWTtnGKSMMs-A0dJxsRi-KSs7--UZiyE83OXeCl8pyI7TbYikmYWUYsLjBtPPycxtz-86yHOeEjR6fGL6ZXDxMh9dsiv8AH6-QsA</recordid><startdate>19920401</startdate><enddate>19920401</enddate><creator>Paoni, Nicholas F.</creator><creator>Refino, Canio J.</creator><creator>Brady, Kevin</creator><creator>Pena, Luis C.</creator><creator>Nguyen, Hung V.</creator><creator>Kerr, Elizabeth M.</creator><creator>Johnson, Adriana C.</creator><creator>Wurm, Florian M.</creator><creator>van Reis, Robert</creator><creator>Botstein, David</creator><creator>Bennett, William F.</creator><general>Oxford University Press</general><scope>BSCLL</scope><scope>IQODW</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>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M81</scope><scope>P64</scope></search><sort><creationdate>19920401</creationdate><title>Involvement of residues 296–299 in the enzymatic activity of tissue-type plasminogen activator</title><author>Paoni, Nicholas F. ; Refino, Canio J. ; Brady, Kevin ; Pena, Luis C. ; Nguyen, Hung V. ; Kerr, Elizabeth M. ; Johnson, Adriana C. ; Wurm, Florian M. ; van Reis, Robert ; Botstein, David ; Bennett, William F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c390t-95db9541038030f0561fb3535c1592bfdb2318b0c1353590f59c8f384cc9bc013</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Alanine - chemistry</topic><topic>Amino Acid Sequence</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Enzymes and enzyme inhibitors</topic><topic>fibrin</topic><topic>Fibrin - metabolism</topic><topic>Fibrinogen - metabolism</topic><topic>Fibrinolysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>man</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>mutagenesis</topic><topic>Mutagenesis, Site-Directed</topic><topic>Oligopeptides - metabolism</topic><topic>Plasma</topic><topic>Plasminogen - metabolism</topic><topic>plasminogen activator</topic><topic>Protein Engineering</topic><topic>site-directed mutagenesis</topic><topic>specificity</topic><topic>Tissue Plasminogen Activator - chemistry</topic><topic>Tissue Plasminogen Activator - metabolism</topic><topic>tissue-type plasminogen activator</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Paoni, Nicholas F.</creatorcontrib><creatorcontrib>Refino, Canio J.</creatorcontrib><creatorcontrib>Brady, Kevin</creatorcontrib><creatorcontrib>Pena, Luis C.</creatorcontrib><creatorcontrib>Nguyen, Hung V.</creatorcontrib><creatorcontrib>Kerr, Elizabeth M.</creatorcontrib><creatorcontrib>Johnson, Adriana C.</creatorcontrib><creatorcontrib>Wurm, Florian M.</creatorcontrib><creatorcontrib>van Reis, Robert</creatorcontrib><creatorcontrib>Botstein, David</creatorcontrib><creatorcontrib>Bennett, William F.</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Protein engineering</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Paoni, Nicholas F.</au><au>Refino, Canio J.</au><au>Brady, Kevin</au><au>Pena, Luis C.</au><au>Nguyen, Hung V.</au><au>Kerr, Elizabeth M.</au><au>Johnson, Adriana C.</au><au>Wurm, Florian M.</au><au>van Reis, Robert</au><au>Botstein, David</au><au>Bennett, William F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Involvement of residues 296–299 in the enzymatic activity of tissue-type plasminogen activator</atitle><jtitle>Protein engineering</jtitle><addtitle>Protein Eng</addtitle><date>1992-04-01</date><risdate>1992</risdate><volume>5</volume><issue>3</issue><spage>259</spage><epage>266</epage><pages>259-266</pages><issn>1741-0126</issn><issn>0269-2139</issn><eissn>1741-0134</eissn><eissn>1460-213X</eissn><coden>PRENE9</coden><abstract>The tetra-alanine substitution variant KHRR 296–299 AAAA of tissue-type plasminogen activator (t-PA) was previously shown to have enhanced fibrin specificity and enhanced activity in the presence of fibrin compared with the wild-type form of the molecule. The structural requirements for these alterations in enzymatic activity were investigated by constructing several amino acid substitution variants at each of the positions from 296 to 299 and evaluating their activities under a variety of conditions. Effects on plasminogen activator activity were common among the point mutants at positions 296–299; nearly all had a phenotype similar to the KHRR 296–299 AAAA variant. The greatest effects on enzymatic function were found with multiple substitution variants, but some single charge reversals and proline substitutions had substantial effects. The enhanced fibrin specificity of KHRR 296–299 AAAA t-PA results in less fibrinogenolysis than seen with wild-type t-PA. Approximately four times greater concentration of KHRR 296–299 AAAA compared with wild-type t-PA was required to consume 50% of the fibrinogen in human plasma.</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><pmid>1409547</pmid><doi>10.1093/protein/5.3.259</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1741-0126 |
| ispartof | Protein engineering, 1992-04, Vol.5 (3), p.259-266 |
| issn | 1741-0126 0269-2139 1741-0134 1460-213X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16501377 |
| source | MEDLINE; Alma/SFX Local Collection; Oxford University Press Journals Digital Archive Legacy |
| subjects | Alanine - chemistry Amino Acid Sequence Analytical, structural and metabolic biochemistry Biological and medical sciences Enzymes and enzyme inhibitors fibrin Fibrin - metabolism Fibrinogen - metabolism Fibrinolysis Fundamental and applied biological sciences. Psychology man Models, Molecular Molecular Sequence Data mutagenesis Mutagenesis, Site-Directed Oligopeptides - metabolism Plasma Plasminogen - metabolism plasminogen activator Protein Engineering site-directed mutagenesis specificity Tissue Plasminogen Activator - chemistry Tissue Plasminogen Activator - metabolism tissue-type plasminogen activator |
| title | Involvement of residues 296–299 in the enzymatic activity of tissue-type plasminogen activator |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-16T04%3A05%3A27IST&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=Involvement%20of%20residues%20296%E2%80%93299%20in%20the%20enzymatic%20activity%20of%20tissue-type%20plasminogen%20activator&rft.jtitle=Protein%20engineering&rft.au=Paoni,%20Nicholas%20F.&rft.date=1992-04-01&rft.volume=5&rft.issue=3&rft.spage=259&rft.epage=266&rft.pages=259-266&rft.issn=1741-0126&rft.eissn=1741-0134&rft.coden=PRENE9&rft_id=info:doi/10.1093/protein/5.3.259&rft_dat=%3Cproquest_cross%3E16501377%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=16501377&rft_id=info:pmid/1409547&rfr_iscdi=true |