{"id":"https://openalex.org/W2940112946","doi":"https://doi.org/10.2144/btn-2018-0135","title":"A Robust Fractionation Method for Protein Subcellular Localization Studies in Escherichia Coli","display_name":"A Robust Fractionation Method for Protein Subcellular Localization Studies in Escherichia Coli","publication_year":2019,"publication_date":"2019-04-01","ids":{"openalex":"https://openalex.org/W2940112946","doi":"https://doi.org/10.2144/btn-2018-0135","mag":"2940112946","pmid":"https://pubmed.ncbi.nlm.nih.gov/30987443"},"language":"en","primary_location":{"id":"doi:10.2144/btn-2018-0135","doi":"https://doi.org/10.2144/btn-2018-0135","is_oa":true,"landing_page_url":"https://doi.org/10.2144/btn-2018-0135","pdf_url":"https://www.future-science.com/doi/pdf/10.2144/btn-2018-0135","source":{"id":"https://openalex.org/S79990748","display_name":"BioTechniques","issn_l":"0736-6205","issn":["0736-6205","1940-9818"],"is_oa":true,"is_in_doaj":true,"is_core":true,"listed_in":["cwts-core","doaj","dongbi-c","jufo-1","medline","norway-1","poland-70","russia-white-list-2","vabb-shw"],"host_organization":"https://openalex.org/P4310320316","host_organization_name":"Future Science Ltd","host_organization_lineage":["https://openalex.org/P4310320316"],"host_organization_lineage_names":["Future Science Ltd"],"type":"journal"},"license":"cc-by-nc-nd","license_id":"https://openalex.org/licenses/cc-by-nc-nd","version":"publishedVersion","is_accepted":true,"is_published":true,"raw_source_name":"BioTechniques","raw_type":"journal-article"},"volume":null,"type":"article","indexed_in":["crossref","doaj","pubmed"],"open_access":{"is_oa":true,"oa_status":"gold","oa_url":"https://www.future-science.com/doi/pdf/10.2144/btn-2018-0135","any_repository_has_fulltext":true},"authorships":[{"author_position":"first","author":{"id":null,"display_name":"Gilles Malherbe","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I20581793","display_name":"University of Kent","ror":"https://ror.org/00xkeyj56","country_code":"GB","type":"education","lineage":["https://openalex.org/I20581793"]},{"id":"https://openalex.org/I4210134054","display_name":"UCB Pharma (United Kingdom)","ror":"https://ror.org/03428qp74","country_code":"GB","type":"company","lineage":["https://openalex.org/I4210134054","https://openalex.org/I85722119"]}],"countries":["GB"],"is_corresponding":false,"raw_author_name":"Gilles Malherbe","raw_affiliation_strings":["School of Biosciences, University of Kent, Canterbury, UK","UCB Celltech, Slough, UK","UCB Celltech, Slough, UK School of Biosciences, University of Kent, Canterbury, UK"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"School of Biosciences, University of Kent, Canterbury, UK","institution_ids":["https://openalex.org/I20581793"]},{"raw_affiliation_string":"UCB Celltech, Slough, UK","institution_ids":["https://openalex.org/I4210134054"]},{"raw_affiliation_string":"UCB Celltech, Slough, UK School of Biosciences, University of Kent, Canterbury, UK","institution_ids":["https://openalex.org/I20581793"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5070702643","display_name":"David P. Humphreys","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I4210134054","display_name":"UCB Pharma (United Kingdom)","ror":"https://ror.org/03428qp74","country_code":"GB","type":"company","lineage":["https://openalex.org/I4210134054","https://openalex.org/I85722119"]}],"countries":["GB"],"is_corresponding":false,"raw_author_name":"David Paul Humphreys","raw_affiliation_strings":["UCB Celltech, Slough, UK"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"UCB Celltech, Slough, UK","institution_ids":["https://openalex.org/I4210134054"]}]},{"author_position":"last","author":{"id":"https://openalex.org/A5038586187","display_name":"Emma Dav\u00e9","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I4210134054","display_name":"UCB Pharma (United Kingdom)","ror":"https://ror.org/03428qp74","country_code":"GB","type":"company","lineage":["https://openalex.org/I4210134054","https://openalex.org/I85722119"]}],"countries":["GB"],"is_corresponding":false,"raw_author_name":"Emma Dav\u00e9","raw_affiliation_strings":["UCB Celltech, Slough, UK"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"UCB Celltech, Slough, UK","institution_ids":["https://openalex.org/I4210134054"]}]}],"institutions":[],"countries_distinct_count":1,"institutions_distinct_count":2,"corresponding_author_ids":[],"corresponding_institution_ids":[],"apc_list":{"value":1415,"currency":"USD","value_usd":1415},"apc_paid":{"value":1415,"currency":"USD","value_usd":1415},"fwci":2.2535,"has_fulltext":true,"cited_by_count":42,"citation_normalized_percentile":{"value":0.87623106,"is_in_top_1_percent":false,"is_in_top_10_percent":false},"cited_by_percentile_year":{"min":93,"max":99},"biblio":{"volume":"66","issue":"4","first_page":"171","last_page":"178"},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T10120","display_name":"Bacterial Genetics and Biotechnology","score":0.7993000149726868,"subfield":{"id":"https://openalex.org/subfields/1311","display_name":"Genetics"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},"topics":[{"id":"https://openalex.org/T10120","display_name":"Bacterial Genetics and Biotechnology","score":0.7993000149726868,"subfield":{"id":"https://openalex.org/subfields/1311","display_name":"Genetics"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T11124","display_name":"Protein purification and stability","score":0.163100004196167,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T10519","display_name":"Advanced Proteomics Techniques and Applications","score":0.009999999776482582,"subfield":{"id":"https://openalex.org/subfields/1607","display_name":"Spectroscopy"},"field":{"id":"https://openalex.org/fields/16","display_name":"Chemistry"},"domain":{"id":"https://openalex.org/domains/3","display_name":"Physical Sciences"}}],"keywords":[{"id":"https://openalex.org/keywords/subcellular-fractionation","display_name":"subcellular fractionation","score":0.8889999985694885},{"id":"https://openalex.org/keywords/protein-localization","display_name":"protein localization","score":0.8859999775886536},{"id":"https://openalex.org/keywords/escherichia-coli","display_name":"Escherichia coli","score":0.9710000157356262},{"id":"https://openalex.org/keywords/cytosolic-proteins","display_name":"cytosolic proteins","score":0.8149999976158142},{"id":"https://openalex.org/keywords/magnesium","display_name":"magnesium","score":0.42500001192092896},{"id":"https://openalex.org/keywords/sonication","display_name":"sonication","score":0.8769999742507935},{"id":"https://openalex.org/keywords/ultracentrifugation","display_name":"ultracentrifugation","score":0.9909999966621399},{"id":"https://openalex.org/keywords/gram-negative-bacteria","display_name":"Gram-negative bacteria","score":0.7369999885559082}],"concepts":[{"id":"https://openalex.org/C201663137","wikidata":"https://www.wikidata.org/wiki/Q21198990","display_name":"Periplasmic space","level":4,"score":0.9652383923530579},{"id":"https://openalex.org/C27239569","wikidata":"https://www.wikidata.org/wiki/Q189029","display_name":"Cell fractionation","level":3,"score":0.7952776551246643},{"id":"https://openalex.org/C547475151","wikidata":"https://www.wikidata.org/wiki/Q25419","display_name":"Escherichia coli","level":3,"score":0.7362871170043945},{"id":"https://openalex.org/C190062978","wikidata":"https://www.wikidata.org/wiki/Q79899","display_name":"Cytoplasm","level":2,"score":0.7008078098297119},{"id":"https://openalex.org/C97428945","wikidata":"https://www.wikidata.org/wiki/Q15305109","display_name":"Fractionation","level":2,"score":0.6482667326927185},{"id":"https://openalex.org/C104628117","wikidata":"https://www.wikidata.org/wiki/Q3964828","display_name":"Sonication","level":2,"score":0.5881360173225403},{"id":"https://openalex.org/C26268613","wikidata":"https://www.wikidata.org/wiki/Q187643","display_name":"Cell disruption","level":2,"score":0.5833621025085449},{"id":"https://openalex.org/C57409179","wikidata":"https://www.wikidata.org/wiki/Q1218293","display_name":"Lysis","level":2,"score":0.561227023601532},{"id":"https://openalex.org/C168529131","wikidata":"https://www.wikidata.org/wiki/Q13656562","display_name":"Osmotic shock","level":3,"score":0.5418906807899475},{"id":"https://openalex.org/C2776879804","wikidata":"https://www.wikidata.org/wiki/Q7630919","display_name":"Subcellular localization","level":3,"score":0.49348676204681396},{"id":"https://openalex.org/C55493867","wikidata":"https://www.wikidata.org/wiki/Q7094","display_name":"Biochemistry","level":1,"score":0.4924079179763794},{"id":"https://openalex.org/C54689828","wikidata":"https://www.wikidata.org/wiki/Q2029259","display_name":"Ultracentrifuge","level":2,"score":0.46125921607017517},{"id":"https://openalex.org/C523546767","wikidata":"https://www.wikidata.org/wiki/Q10876","display_name":"Bacteria","level":2,"score":0.45396292209625244},{"id":"https://openalex.org/C140051345","wikidata":"https://www.wikidata.org/wiki/Q7251501","display_name":"Protein subcellular localization prediction","level":3,"score":0.44582757353782654},{"id":"https://openalex.org/C185592680","wikidata":"https://www.wikidata.org/wiki/Q2329","display_name":"Chemistry","level":0,"score":0.4436536133289337},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.3679478168487549},{"id":"https://openalex.org/C43617362","wikidata":"https://www.wikidata.org/wiki/Q170050","display_name":"Chromatography","level":1,"score":0.34123343229293823},{"id":"https://openalex.org/C181199279","wikidata":"https://www.wikidata.org/wiki/Q8047","display_name":"Enzyme","level":2,"score":0.14784416556358337},{"id":"https://openalex.org/C104317684","wikidata":"https://www.wikidata.org/wiki/Q7187","display_name":"Gene","level":2,"score":0.09026062488555908},{"id":"https://openalex.org/C54355233","wikidata":"https://www.wikidata.org/wiki/Q7162","display_name":"Genetics","level":1,"score":0.0}],"mesh":[{"descriptor_ui":"D002458","descriptor_name":"Cell Fractionation","qualifier_ui":"Q000379","qualifier_name":"methods","is_major_topic":true},{"descriptor_ui":"D003080","descriptor_name":"Cold Temperature","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D004926","descriptor_name":"Escherichia coli","qualifier_ui":"Q000166","qualifier_name":"cytology","is_major_topic":true},{"descriptor_ui":"D004926","descriptor_name":"Escherichia coli","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":false},{"descriptor_ui":"D009997","descriptor_name":"Osmotic Pressure","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D011994","descriptor_name":"Recombinant Proteins","qualifier_ui":"Q000032","qualifier_name":"analysis","is_major_topic":true},{"descriptor_ui":"D015153","descriptor_name":"Blotting, Western","qualifier_ui":"Q000379","qualifier_name":"methods","is_major_topic":false},{"descriptor_ui":"D019897","descriptor_name":"Periplasm","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":true},{"descriptor_ui":"D029968","descriptor_name":"Escherichia coli Proteins","qualifier_ui":"Q000032","qualifier_name":"analysis","is_major_topic":true}],"locations_count":3,"locations":[{"id":"doi:10.2144/btn-2018-0135","doi":"https://doi.org/10.2144/btn-2018-0135","is_oa":true,"landing_page_url":"https://doi.org/10.2144/btn-2018-0135","pdf_url":"https://www.future-science.com/doi/pdf/10.2144/btn-2018-0135","source":{"id":"https://openalex.org/S79990748","display_name":"BioTechniques","issn_l":"0736-6205","issn":["0736-6205","1940-9818"],"is_oa":true,"is_in_doaj":true,"is_core":true,"listed_in":["cwts-core","doaj","dongbi-c","jufo-1","medline","norway-1","poland-70","russia-white-list-2","vabb-shw"],"host_organization":"https://openalex.org/P4310320316","host_organization_name":"Future Science Ltd","host_organization_lineage":["https://openalex.org/P4310320316"],"host_organization_lineage_names":["Future Science Ltd"],"type":"journal"},"license":"cc-by-nc-nd","license_id":"https://openalex.org/licenses/cc-by-nc-nd","version":"publishedVersion","is_accepted":true,"is_published":true,"raw_source_name":"BioTechniques","raw_type":"journal-article"},{"id":"pmid:30987443","doi":null,"is_oa":false,"landing_page_url":"https://pubmed.ncbi.nlm.nih.gov/30987443","pdf_url":null,"source":{"id":"https://openalex.org/S4306525036","display_name":"PubMed","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":"https://openalex.org/I1299303238","host_organization_name":"National Institutes of Health","host_organization_lineage":["https://openalex.org/I1299303238"],"host_organization_lineage_names":[],"type":"repository"},"license":null,"license_id":null,"version":"publishedVersion","is_accepted":true,"is_published":true,"raw_source_name":"BioTechniques","raw_type":"Journal Article"},{"id":"pmh:oai:doaj.org/article:172f1e98938c49359f826783c5a43cbe","doi":null,"is_oa":true,"landing_page_url":"https://doaj.org/article/172f1e98938c49359f826783c5a43cbe","pdf_url":null,"source":{"id":"https://openalex.org/S4306401280","display_name":"DOAJ (DOAJ: Directory of Open Access Journals)","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":null,"host_organization_name":null,"host_organization_lineage":[],"host_organization_lineage_names":[],"type":"repository"},"license":"cc-by-sa","license_id":"https://openalex.org/licenses/cc-by-sa","version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":"BioTechniques, Vol 66, Iss 4, Pp 171-178 (2019)","raw_type":"article"}],"best_oa_location":{"id":"doi:10.2144/btn-2018-0135","doi":"https://doi.org/10.2144/btn-2018-0135","is_oa":true,"landing_page_url":"https://doi.org/10.2144/btn-2018-0135","pdf_url":"https://www.future-science.com/doi/pdf/10.2144/btn-2018-0135","source":{"id":"https://openalex.org/S79990748","display_name":"BioTechniques","issn_l":"0736-6205","issn":["0736-6205","1940-9818"],"is_oa":true,"is_in_doaj":true,"is_core":true,"listed_in":["cwts-core","doaj","dongbi-c","jufo-1","medline","norway-1","poland-70","russia-white-list-2","vabb-shw"],"host_organization":"https://openalex.org/P4310320316","host_organization_name":"Future Science Ltd","host_organization_lineage":["https://openalex.org/P4310320316"],"host_organization_lineage_names":["Future Science Ltd"],"type":"journal"},"license":"cc-by-nc-nd","license_id":"https://openalex.org/licenses/cc-by-nc-nd","version":"publishedVersion","is_accepted":true,"is_published":true,"raw_source_name":"BioTechniques","raw_type":"journal-article"},"sustainable_development_goals":[],"sustainable_development_goals_aurora":[],"study_designs":[],"awards":[],"funders":[],"has_content":{"grobid_xml":true,"pdf":true},"content_urls":{"pdf":"https://content.openalex.org/works/W2940112946.pdf","grobid_xml":"https://content.openalex.org/works/W2940112946.grobid-xml"},"referenced_works_count":47,"referenced_works":["https://openalex.org/W155901324","https://openalex.org/W1512810258","https://openalex.org/W1537641994","https://openalex.org/W1833589815","https://openalex.org/W1925087414","https://openalex.org/W1962713699","https://openalex.org/W1965881058","https://openalex.org/W1975920595","https://openalex.org/W1993390189","https://openalex.org/W1996123206","https://openalex.org/W1999986549","https://openalex.org/W2000212768","https://openalex.org/W2010816372","https://openalex.org/W2016667239","https://openalex.org/W2030049548","https://openalex.org/W2044825931","https://openalex.org/W2049748661","https://openalex.org/W2049901993","https://openalex.org/W2055600356","https://openalex.org/W2063968162","https://openalex.org/W2083021096","https://openalex.org/W2097592065","https://openalex.org/W2099091042","https://openalex.org/W2105887681","https://openalex.org/W2107227535","https://openalex.org/W2111198021","https://openalex.org/W2115631722","https://openalex.org/W2135604463","https://openalex.org/W2138820253","https://openalex.org/W2140536590","https://openalex.org/W2142524368","https://openalex.org/W2148054445","https://openalex.org/W2148904014","https://openalex.org/W2150733248","https://openalex.org/W2160382463","https://openalex.org/W2162792752","https://openalex.org/W2170632475","https://openalex.org/W2308145610","https://openalex.org/W2487270267","https://openalex.org/W2520595272","https://openalex.org/W2552158877","https://openalex.org/W2592567520","https://openalex.org/W2609207099","https://openalex.org/W2627092082","https://openalex.org/W2736646038","https://openalex.org/W2779520800","https://openalex.org/W2804007336"],"related_works":["https://openalex.org/W4388422389","https://openalex.org/W2024987906","https://openalex.org/W2030049548","https://openalex.org/W4392779855","https://openalex.org/W2366506100","https://openalex.org/W6946343533","https://openalex.org/W2175663749","https://openalex.org/W2729122423","https://openalex.org/W2362292877","https://openalex.org/W2415329421"],"abstract_inverted_index":{"Fractionation":[0],"in":[1,13,51,114,123],"Gram-negative":[2],"bacteria":[3],"is":[4],"used":[5],"to":[6,54,98,102],"identify":[7],"the":[8,15,104],"subcellular":[9],"localization":[10,16],"of":[11,17,23,83,120],"proteins,":[12],"particular":[14],"exported":[18],"recombinant":[19],"proteins.":[20,86],"The":[21,87],"process":[22,57,88],"cell":[24,48],"fractionation":[25],"can":[26],"be":[27],"fraught":[28],"with":[29,95],"cross-contamination":[30],"issues":[31],"and":[32,46,79,100,117],"often":[33],"lacks":[34],"supporting":[35],"data":[36],"for":[37],"fraction":[38],"purity.":[39],"Here,":[40],"we":[41],"compare":[42],"three":[43],"periplasm":[44,90],"extraction":[45],"two":[47],"disruption":[49],"techniques":[50],"different":[52],"combinations":[53],"investigate":[55],"which":[56],"gives":[58,75],"uncontaminated":[59],"compartments":[60],"from":[61,106],"Escherichia":[62],"coli.":[63],"From":[64],"these":[65],"data,":[66],"a":[67,80],"robust":[68],"method":[69,110],"named":[70],"PureFrac":[71],"was":[72],"compiled":[73],"that":[74],"pure":[76],"periplasmic":[77],"fractions":[78],"superior":[81],"recovery":[82],"soluble":[84],"cytoplasmic":[85],"extracts":[89],"using":[91],"cold":[92],"osmotic":[93],"shock":[94],"magnesium,":[96],"prior":[97],"sonication":[99],"ultracentrifugation":[101],"separate":[103],"cytoplasm":[105],"insoluble":[107],"material.":[108],"This":[109],"handles":[111],"cells":[112],"cultivated":[113],"various":[115],"conditions":[116],"allows":[118],"preparation":[119],"active":[121],"proteins":[122],"their":[124],"respective":[125],"compartments.":[126]},"counts_by_year":[{"year":2026,"cited_by_count":4},{"year":2025,"cited_by_count":5},{"year":2024,"cited_by_count":6},{"year":2023,"cited_by_count":12},{"year":2022,"cited_by_count":9},{"year":2021,"cited_by_count":2},{"year":2020,"cited_by_count":4}],"updated_date":"2026-10-31T12:09:11.096373","created_date":"2019-04-25T00:00:00"}