{"corpus_id":2649209,"paper_sha":"3a724e68c44be2c4aec16ea22bad23d626a049bc","doi":"10.1371/journal.pone.0008119","arxiv_id":null,"pmid":19956581,"pmcid":"2780291","mag_id":2097715383,"dblp_id":null,"acl_id":null,"title":"Simplified, Enhanced Protein Purification Using an Inducible, Autoprocessing Enzyme Tag","year":2009,"publication_date":"2009-12-02","venue":"PLoS ONE","journal":{"name":"PLoS ONE","pages":null,"volume":"4"},"journal_issn":null,"journal_title":null,"publication_types":["JournalArticle"],"pubmed_pub_types":["Journal Article","Research Support, N.I.H., Extramural","Research Support, Non-U.S. Gov't"],"s2_fields_of_study":["Biology","Medicine","Chemistry"],"reference_count":23,"citation_count":92,"influential_citation_count":4,"is_open_access":true,"arxiv_categories":null,"arxiv_license":null,"arxiv_journal_ref":null,"mesh_headings":[{"d":"Animals","mj":false,"ui":"D000818"},{"d":"Biochemistry","mj":false,"qs":[{"q":"methods","mj":true,"ui":"Q000379"}],"ui":"D001671"},{"d":"Coenzyme A Ligases","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D003066"},{"d":"Genetic Vectors","mj":false,"qs":[{"q":"genetics","mj":false,"ui":"Q000235"}],"ui":"D005822"},{"d":"Histidine","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D006639"},{"d":"Matrix Metalloproteinase 12","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D053508"},{"d":"Mice","mj":false,"ui":"D051379"},{"d":"Oligopeptides","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D009842"},{"d":"Peptide Hydrolases","mj":false,"qs":[{"q":"metabolism","mj":true,"ui":"Q000378"}],"ui":"D010447"},{"d":"Protein Stability","mj":false,"ui":"D055550"},{"d":"Protein Structure, Tertiary","mj":false,"ui":"D017434"},{"d":"Protozoan Proteins","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D015800"},{"d":"Recombinant Fusion Proteins","mj":false,"qs":[{"q":"chemistry","mj":false,"ui":"Q000737"},{"q":"isolation & purification","mj":true,"ui":"Q000302"},{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D011993"},{"d":"Solubility","mj":false,"ui":"D012995"}],"chemicals":[{"n":"His-His-His-His-His-His","ui":"C471213","reg":"0"},{"n":"Oligopeptides","ui":"D009842","reg":"0"},{"n":"Protozoan Proteins","ui":"D015800","reg":"0"},{"n":"Recombinant Fusion Proteins","ui":"D011993","reg":"0"},{"n":"Histidine","ui":"D006639","reg":"4QD397987E"},{"n":"Peptide Hydrolases","ui":"D010447","reg":"EC 3.4.-"},{"n":"Matrix Metalloproteinase 12","ui":"D053508","reg":"EC 3.4.24.65"},{"n":"Coenzyme A Ligases","ui":"D003066","reg":"EC 6.2.1.-"},{"n":"biotin-CoA ligase","ui":"C051127","reg":"EC 6.2.1.11"}],"comments_corrections":null,"source_flags":5,"s2_open_access_pdf_url":"https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0008119&type=printable","s2_open_access_landing_url":"https://www.semanticscholar.org/paper/3a724e68c44be2c4aec16ea22bad23d626a049bc","s2_open_access_license":"CCBY","s2_open_access_status":"GOLD","pmc_open_access_pdf_url":null,"pmc_open_access_landing_url":null,"pmc_open_access_license":null,"pmc_open_access_status":null,"unpaywall_open_access_pdf_url":null,"unpaywall_open_access_landing_url":null,"unpaywall_open_access_license":null,"unpaywall_open_access_status":null,"abstract":"We introduce a new method for purifying recombinant proteins expressed in bacteria using a highly specific, inducible, self-cleaving protease tag. 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This method condenses affinity chromatography and fusion tag cleavage into a single step, obviating the need for exogenous protease addition to remove the fusion tag(s) and increasing the efficiency of tag separation. Furthermore, in addition to being timesaving, versatile, and inexpensive, our results indicate that the CPD purification system can enhance the expression, integrity, and solubility of intractable proteins from diverse organisms.","claims":[{"public_id":"cl_eb19fbe66694be687565bc6206aa0be5","status":"active","text":"A highly specific, inducible, self-cleaving protease tag enables purification of recombinant proteins expressed in bacteria.","confidence":0.97,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_eb19fbe66694be687565bc6206aa0be5"},{"public_id":"cl_ff3b37d3c1039ace14670cfdfd74674a","status":"active","text":"His6-tagged CPD fused to the C-terminus of target proteins can be purified from Escherichia coli lysates by metal ion affinity chromatography and then cleaved on-column by addition of InsP6 to release untagged target 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