Fine tuning of spatial arrangement of enzymes in a PCNA-mediated multienzyme complex using a rigid poly-L-proline linker
Inspired by natural multienzyme complexes, many types of artificial multienzyme complexes have recently been constructed. We previously constructed a self-assembled complex of a bacterial cytochrome P450 and its ferredoxin and ferredoxin reductase partners using heterotrimerization of proliferating...
Gespeichert in:
| Veröffentlicht in: | PloS one 2013-09, Vol.8 (9), p.e75114-e75114 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | e75114 |
|---|---|
| container_issue | 9 |
| container_start_page | e75114 |
| container_title | PloS one |
| container_volume | 8 |
| creator | Haga, Tomoaki Hirakawa, Hidehiko Nagamune, Teruyuki |
| description | Inspired by natural multienzyme complexes, many types of artificial multienzyme complexes have recently been constructed. We previously constructed a self-assembled complex of a bacterial cytochrome P450 and its ferredoxin and ferredoxin reductase partners using heterotrimerization of proliferating cell nuclear antigen (PCNA) from Sulfolobus solfataricus. In this study, we inserted different peptide linkers between ferredoxin and the PCNA subunit, and examined the effect on activity of the self-assembled multienzyme complex. Although the activity was affected by the lengths of both the rigid poly-L-proline-rich linkers and the flexible Gly4-Ser repeating linkers, the poly-L-proline-rich linkers provided the greatest activity enhancement. The optimized poly-L-proline-rich linker enhanced the activity 1.9-fold compared with the GGGGSLVPRGSGGGGS linker used in the previously reported complex, while the Gly4-Ser repeating linkers, (G4S)n (n = 1-6), did not yield higher activity than the maximum activity by the optimized poly-L-proline linker. Both the rigidity/flexibility and length of the peptide linker were found to be important for enhancing the overall activity of the multienzyme complex. |
| doi_str_mv | 10.1371/journal.pone.0075114 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_1430457356</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A478415952</galeid><doaj_id>oai_doaj_org_article_1715ff768bca49f1bc16d5f2cf07bb5f</doaj_id><sourcerecordid>A478415952</sourcerecordid><originalsourceid>FETCH-LOGICAL-c758t-21ff8f85df81ef3682f27d74d3861f1592bff4120e12bca0c12d2188926c55063</originalsourceid><addsrcrecordid>eNqNk11v0zAUhiMEYmPwDxBEQkJwkeLPOL1BqioGlSqG-Lq1XMdOXRw7sxPU8utx1m5q0S5QpMQ6fs57cl77ZNlzCCYQM_hu44fghJ103qkJAIxCSB5k53CKUVEigB8erc-yJzFuAKC4KsvH2RkigAA8RefZ9tI4lfeDM67Jvc5jJ3ojbC5CEK5RrXL9GFbuz65VMTcuF_mX-edZ0araiF7VeTvY3uz3c-nbzqptPsRRTuTBNKbOO293xbLogrdjsfT6pcLT7JEWNqpnh-9F9uPyw_f5p2J59XExny0LyWjVFwhqXemK1rqCSuOyQhqxmpE6dQI1pFO00ppABBREKymAhKhGsKqmqJSUghJfZC_3up31kR9MixwSDAhlmI7EYk_UXmx4F0wrwo57YfhNwIeGi9AbaRWHDFKtWVmlUmSq4UrCsqYaSQ3YakV10np_qDaskkMy2ReEPRE93XFmzRv_m2NWEshIEnhzEAj-elCx562JUlkrnPLDzX9jxDBBVUJf_YPe392BakRqwDjtU105ivIZYRVJFlKUqMk9VHpq1RqZbpg2KX6S8PYkITG92vaNGGLki29f_5-9-nnKvj5i10rYfh29HXrjXTwFyR6UwccYlL4zGQI-DsitG3wcEH4YkJT24viA7pJuJwL_BevDC24</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1430457356</pqid></control><display><type>article</type><title>Fine tuning of spatial arrangement of enzymes in a PCNA-mediated multienzyme complex using a rigid poly-L-proline linker</title><source>Public Library of Science (PLoS) Journals Open Access</source><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Haga, Tomoaki ; Hirakawa, Hidehiko ; Nagamune, Teruyuki</creator><contributor>Isalan, Mark</contributor><creatorcontrib>Haga, Tomoaki ; Hirakawa, Hidehiko ; Nagamune, Teruyuki ; Isalan, Mark</creatorcontrib><description>Inspired by natural multienzyme complexes, many types of artificial multienzyme complexes have recently been constructed. We previously constructed a self-assembled complex of a bacterial cytochrome P450 and its ferredoxin and ferredoxin reductase partners using heterotrimerization of proliferating cell nuclear antigen (PCNA) from Sulfolobus solfataricus. In this study, we inserted different peptide linkers between ferredoxin and the PCNA subunit, and examined the effect on activity of the self-assembled multienzyme complex. Although the activity was affected by the lengths of both the rigid poly-L-proline-rich linkers and the flexible Gly4-Ser repeating linkers, the poly-L-proline-rich linkers provided the greatest activity enhancement. The optimized poly-L-proline-rich linker enhanced the activity 1.9-fold compared with the GGGGSLVPRGSGGGGS linker used in the previously reported complex, while the Gly4-Ser repeating linkers, (G4S)n (n = 1-6), did not yield higher activity than the maximum activity by the optimized poly-L-proline linker. Both the rigidity/flexibility and length of the peptide linker were found to be important for enhancing the overall activity of the multienzyme complex.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0075114</identifier><identifier>PMID: 24040392</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Amino Acid Sequence ; Bacteria ; Biochemistry ; Biotechnology ; Camphor 5-Monooxygenase - chemistry ; Chemistry ; Cytochrome ; Cytochrome P450 ; Cytochromes c - metabolism ; Deoxyribonucleic acid ; DNA ; Engineering schools ; Enzymes ; Ferredoxin ; Ferredoxin reductase ; Ferredoxins - chemistry ; L-Proline ; Models, Molecular ; Molecular Sequence Data ; Multienzyme complexes ; Multienzyme Complexes - chemistry ; Multienzyme Complexes - genetics ; Multienzyme Complexes - metabolism ; Oxidation-Reduction ; Peptides ; Peptides - metabolism ; Plasmids ; Potassium ; Proliferating cell nuclear antigen ; Proliferating Cell Nuclear Antigen - chemistry ; Proliferating Cell Nuclear Antigen - metabolism ; Proline ; Protein Engineering - methods ; Protein Multimerization ; Protein Structure, Quaternary ; Proteins ; Recombinant Fusion Proteins - chemistry ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Rigidity ; Self assembly ; Structure-Activity Relationship</subject><ispartof>PloS one, 2013-09, Vol.8 (9), p.e75114-e75114</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Haga et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Haga et al 2013 Haga et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-21ff8f85df81ef3682f27d74d3861f1592bff4120e12bca0c12d2188926c55063</citedby><cites>FETCH-LOGICAL-c758t-21ff8f85df81ef3682f27d74d3861f1592bff4120e12bca0c12d2188926c55063</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3764174/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3764174/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24040392$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Isalan, Mark</contributor><creatorcontrib>Haga, Tomoaki</creatorcontrib><creatorcontrib>Hirakawa, Hidehiko</creatorcontrib><creatorcontrib>Nagamune, Teruyuki</creatorcontrib><title>Fine tuning of spatial arrangement of enzymes in a PCNA-mediated multienzyme complex using a rigid poly-L-proline linker</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Inspired by natural multienzyme complexes, many types of artificial multienzyme complexes have recently been constructed. We previously constructed a self-assembled complex of a bacterial cytochrome P450 and its ferredoxin and ferredoxin reductase partners using heterotrimerization of proliferating cell nuclear antigen (PCNA) from Sulfolobus solfataricus. In this study, we inserted different peptide linkers between ferredoxin and the PCNA subunit, and examined the effect on activity of the self-assembled multienzyme complex. Although the activity was affected by the lengths of both the rigid poly-L-proline-rich linkers and the flexible Gly4-Ser repeating linkers, the poly-L-proline-rich linkers provided the greatest activity enhancement. The optimized poly-L-proline-rich linker enhanced the activity 1.9-fold compared with the GGGGSLVPRGSGGGGS linker used in the previously reported complex, while the Gly4-Ser repeating linkers, (G4S)n (n = 1-6), did not yield higher activity than the maximum activity by the optimized poly-L-proline linker. Both the rigidity/flexibility and length of the peptide linker were found to be important for enhancing the overall activity of the multienzyme complex.</description><subject>Amino Acid Sequence</subject><subject>Bacteria</subject><subject>Biochemistry</subject><subject>Biotechnology</subject><subject>Camphor 5-Monooxygenase - chemistry</subject><subject>Chemistry</subject><subject>Cytochrome</subject><subject>Cytochrome P450</subject><subject>Cytochromes c - metabolism</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Engineering schools</subject><subject>Enzymes</subject><subject>Ferredoxin</subject><subject>Ferredoxin reductase</subject><subject>Ferredoxins - chemistry</subject><subject>L-Proline</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Multienzyme complexes</subject><subject>Multienzyme Complexes - chemistry</subject><subject>Multienzyme Complexes - genetics</subject><subject>Multienzyme Complexes - metabolism</subject><subject>Oxidation-Reduction</subject><subject>Peptides</subject><subject>Peptides - metabolism</subject><subject>Plasmids</subject><subject>Potassium</subject><subject>Proliferating cell nuclear antigen</subject><subject>Proliferating Cell Nuclear Antigen - chemistry</subject><subject>Proliferating Cell Nuclear Antigen - metabolism</subject><subject>Proline</subject><subject>Protein Engineering - methods</subject><subject>Protein Multimerization</subject><subject>Protein Structure, Quaternary</subject><subject>Proteins</subject><subject>Recombinant Fusion Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Rigidity</subject><subject>Self assembly</subject><subject>Structure-Activity Relationship</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk11v0zAUhiMEYmPwDxBEQkJwkeLPOL1BqioGlSqG-Lq1XMdOXRw7sxPU8utx1m5q0S5QpMQ6fs57cl77ZNlzCCYQM_hu44fghJ103qkJAIxCSB5k53CKUVEigB8erc-yJzFuAKC4KsvH2RkigAA8RefZ9tI4lfeDM67Jvc5jJ3ojbC5CEK5RrXL9GFbuz65VMTcuF_mX-edZ0araiF7VeTvY3uz3c-nbzqptPsRRTuTBNKbOO293xbLogrdjsfT6pcLT7JEWNqpnh-9F9uPyw_f5p2J59XExny0LyWjVFwhqXemK1rqCSuOyQhqxmpE6dQI1pFO00ppABBREKymAhKhGsKqmqJSUghJfZC_3up31kR9MixwSDAhlmI7EYk_UXmx4F0wrwo57YfhNwIeGi9AbaRWHDFKtWVmlUmSq4UrCsqYaSQ3YakV10np_qDaskkMy2ReEPRE93XFmzRv_m2NWEshIEnhzEAj-elCx562JUlkrnPLDzX9jxDBBVUJf_YPe392BakRqwDjtU105ivIZYRVJFlKUqMk9VHpq1RqZbpg2KX6S8PYkITG92vaNGGLki29f_5-9-nnKvj5i10rYfh29HXrjXTwFyR6UwccYlL4zGQI-DsitG3wcEH4YkJT24viA7pJuJwL_BevDC24</recordid><startdate>20130905</startdate><enddate>20130905</enddate><creator>Haga, Tomoaki</creator><creator>Hirakawa, Hidehiko</creator><creator>Nagamune, Teruyuki</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130905</creationdate><title>Fine tuning of spatial arrangement of enzymes in a PCNA-mediated multienzyme complex using a rigid poly-L-proline linker</title><author>Haga, Tomoaki ; Hirakawa, Hidehiko ; Nagamune, Teruyuki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-21ff8f85df81ef3682f27d74d3861f1592bff4120e12bca0c12d2188926c55063</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Amino Acid Sequence</topic><topic>Bacteria</topic><topic>Biochemistry</topic><topic>Biotechnology</topic><topic>Camphor 5-Monooxygenase - chemistry</topic><topic>Chemistry</topic><topic>Cytochrome</topic><topic>Cytochrome P450</topic><topic>Cytochromes c - metabolism</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>Engineering schools</topic><topic>Enzymes</topic><topic>Ferredoxin</topic><topic>Ferredoxin reductase</topic><topic>Ferredoxins - chemistry</topic><topic>L-Proline</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Multienzyme complexes</topic><topic>Multienzyme Complexes - chemistry</topic><topic>Multienzyme Complexes - genetics</topic><topic>Multienzyme Complexes - metabolism</topic><topic>Oxidation-Reduction</topic><topic>Peptides</topic><topic>Peptides - metabolism</topic><topic>Plasmids</topic><topic>Potassium</topic><topic>Proliferating cell nuclear antigen</topic><topic>Proliferating Cell Nuclear Antigen - chemistry</topic><topic>Proliferating Cell Nuclear Antigen - metabolism</topic><topic>Proline</topic><topic>Protein Engineering - methods</topic><topic>Protein Multimerization</topic><topic>Protein Structure, Quaternary</topic><topic>Proteins</topic><topic>Recombinant Fusion Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Rigidity</topic><topic>Self assembly</topic><topic>Structure-Activity Relationship</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Haga, Tomoaki</creatorcontrib><creatorcontrib>Hirakawa, Hidehiko</creatorcontrib><creatorcontrib>Nagamune, Teruyuki</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Opposing Viewpoints in Context (Gale)</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Proquest Nursing & Allied Health Source</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Haga, Tomoaki</au><au>Hirakawa, Hidehiko</au><au>Nagamune, Teruyuki</au><au>Isalan, Mark</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fine tuning of spatial arrangement of enzymes in a PCNA-mediated multienzyme complex using a rigid poly-L-proline linker</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-09-05</date><risdate>2013</risdate><volume>8</volume><issue>9</issue><spage>e75114</spage><epage>e75114</epage><pages>e75114-e75114</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Inspired by natural multienzyme complexes, many types of artificial multienzyme complexes have recently been constructed. We previously constructed a self-assembled complex of a bacterial cytochrome P450 and its ferredoxin and ferredoxin reductase partners using heterotrimerization of proliferating cell nuclear antigen (PCNA) from Sulfolobus solfataricus. In this study, we inserted different peptide linkers between ferredoxin and the PCNA subunit, and examined the effect on activity of the self-assembled multienzyme complex. Although the activity was affected by the lengths of both the rigid poly-L-proline-rich linkers and the flexible Gly4-Ser repeating linkers, the poly-L-proline-rich linkers provided the greatest activity enhancement. The optimized poly-L-proline-rich linker enhanced the activity 1.9-fold compared with the GGGGSLVPRGSGGGGS linker used in the previously reported complex, while the Gly4-Ser repeating linkers, (G4S)n (n = 1-6), did not yield higher activity than the maximum activity by the optimized poly-L-proline linker. Both the rigidity/flexibility and length of the peptide linker were found to be important for enhancing the overall activity of the multienzyme complex.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24040392</pmid><doi>10.1371/journal.pone.0075114</doi><tpages>e75114</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2013-09, Vol.8 (9), p.e75114-e75114 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1430457356 |
| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Amino Acid Sequence Bacteria Biochemistry Biotechnology Camphor 5-Monooxygenase - chemistry Chemistry Cytochrome Cytochrome P450 Cytochromes c - metabolism Deoxyribonucleic acid DNA Engineering schools Enzymes Ferredoxin Ferredoxin reductase Ferredoxins - chemistry L-Proline Models, Molecular Molecular Sequence Data Multienzyme complexes Multienzyme Complexes - chemistry Multienzyme Complexes - genetics Multienzyme Complexes - metabolism Oxidation-Reduction Peptides Peptides - metabolism Plasmids Potassium Proliferating cell nuclear antigen Proliferating Cell Nuclear Antigen - chemistry Proliferating Cell Nuclear Antigen - metabolism Proline Protein Engineering - methods Protein Multimerization Protein Structure, Quaternary Proteins Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Rigidity Self assembly Structure-Activity Relationship |
| title | Fine tuning of spatial arrangement of enzymes in a PCNA-mediated multienzyme complex using a rigid poly-L-proline linker |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-29T04%3A02%3A23IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Fine%20tuning%20of%20spatial%20arrangement%20of%20enzymes%20in%20a%20PCNA-mediated%20multienzyme%20complex%20using%20a%20rigid%20poly-L-proline%20linker&rft.jtitle=PloS%20one&rft.au=Haga,%20Tomoaki&rft.date=2013-09-05&rft.volume=8&rft.issue=9&rft.spage=e75114&rft.epage=e75114&rft.pages=e75114-e75114&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0075114&rft_dat=%3Cgale_plos_%3EA478415952%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1430457356&rft_id=info:pmid/24040392&rft_galeid=A478415952&rft_doaj_id=oai_doaj_org_article_1715ff768bca49f1bc16d5f2cf07bb5f&rfr_iscdi=true |