{"id":"https://openalex.org/W4309166196","doi":"https://doi.org/10.1021/acs.bioconjchem.2c00485","title":"Design, Heterologous Expression, and Application of an Immobilized Protein Kinase","display_name":"Design, Heterologous Expression, and Application of an Immobilized Protein Kinase","publication_year":2022,"publication_date":"2022-11-15","ids":{"openalex":"https://openalex.org/W4309166196","doi":"https://doi.org/10.1021/acs.bioconjchem.2c00485","pmid":"https://pubmed.ncbi.nlm.nih.gov/36379001"},"language":"en","primary_location":{"id":"doi:10.1021/acs.bioconjchem.2c00485","doi":"https://doi.org/10.1021/acs.bioconjchem.2c00485","is_oa":false,"landing_page_url":"https://doi.org/10.1021/acs.bioconjchem.2c00485","pdf_url":null,"source":{"id":"https://openalex.org/S155115410","display_name":"Bioconjugate Chemistry","issn_l":"1043-1802","issn":["1043-1802","1520-4812"],"is_oa":false,"is_in_doaj":false,"is_core":true,"listed_in":["cwts-core","dongbi-b","doyens","jufo-1","ki-jl-1","medline","norway-1","poland-100","russia-white-list-1"],"host_organization":"https://openalex.org/P4310320006","host_organization_name":"American Chemical Society","host_organization_lineage":["https://openalex.org/P4310320006"],"host_organization_lineage_names":["American Chemical Society"],"type":"journal"},"license":null,"license_id":null,"version":"publishedVersion","is_accepted":true,"is_published":true,"raw_source_name":"Bioconjugate Chemistry","raw_type":"journal-article"},"volume":null,"type":"article","indexed_in":["crossref","pubmed"],"open_access":{"is_oa":true,"oa_status":"green","oa_url":"https://chemrxiv.org/engage/api-gateway/chemrxiv/assets/orp/resource/item/624326eaed3780eb0d469f08/original/design-and-application-of-an-immobilized-protein-kinase.pdf","any_repository_has_fulltext":true},"authorships":[{"author_position":"first","author":{"id":"https://openalex.org/A5054663256","display_name":"Dalton T. Deane","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I186335123","display_name":"East Carolina University","ror":"https://ror.org/01vx35703","country_code":"US","type":"education","lineage":["https://openalex.org/I186335123"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Dalton T. Deane","raw_affiliation_strings":["Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States","institution_ids":["https://openalex.org/I186335123"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5018895928","display_name":"Thomas A. Cope","orcid":"https://orcid.org/0009-0000-4579-5861","observed_orcids":["https://orcid.org/0009-0000-4579-5861"]},"institutions":[{"id":"https://openalex.org/I17301866","display_name":"University of Alabama","ror":"https://ror.org/03xrrjk67","country_code":"US","type":"education","lineage":["https://openalex.org/I17301866"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Thomas A. Cope","raw_affiliation_strings":["University of Alabama, Tuscaloosa, Alabama 35487, United States"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"University of Alabama, Tuscaloosa, Alabama 35487, United States","institution_ids":["https://openalex.org/I17301866"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5102883367","display_name":"Anna M. Schulz","orcid":"https://orcid.org/0000-0001-6550-7206","observed_orcids":["https://orcid.org/0000-0001-6550-7206"]},"institutions":[{"id":"https://openalex.org/I186335123","display_name":"East Carolina University","ror":"https://ror.org/01vx35703","country_code":"US","type":"education","lineage":["https://openalex.org/I186335123"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Anna M. Schulz","raw_affiliation_strings":["Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States","institution_ids":["https://openalex.org/I186335123"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5043244152","display_name":"Edward T. Bennett","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I114027177","display_name":"University of North Carolina at Chapel Hill","ror":"https://ror.org/0130frc33","country_code":"US","type":"education","lineage":["https://openalex.org/I114027177"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Edward T. Bennett","raw_affiliation_strings":["University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27514, United States"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27514, United States","institution_ids":["https://openalex.org/I114027177"]}]},{"author_position":"last","author":{"id":"https://openalex.org/A5015871731","display_name":"Robert M. Hughes","orcid":"https://orcid.org/0000-0002-4465-4212","observed_orcids":["https://orcid.org/0000-0002-4465-4212"]},"institutions":[{"id":"https://openalex.org/I186335123","display_name":"East Carolina University","ror":"https://ror.org/01vx35703","country_code":"US","type":"education","lineage":["https://openalex.org/I186335123"]}],"countries":["US"],"is_corresponding":true,"raw_author_name":"Robert M. Hughes","raw_affiliation_strings":["Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States"],"raw_orcid":"https://orcid.org/0000-0002-4465-4212","affiliations":[{"raw_affiliation_string":"Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States","institution_ids":["https://openalex.org/I186335123"]}]}],"institutions":[],"countries_distinct_count":1,"institutions_distinct_count":3,"corresponding_author_ids":["https://openalex.org/A5015871731"],"corresponding_institution_ids":["https://openalex.org/I186335123"],"apc_list":{"value":5000,"currency":"USD","value_usd":5000},"apc_paid":null,"fwci":0.1632,"has_fulltext":false,"cited_by_count":4,"citation_normalized_percentile":{"value":0.37478556,"is_in_top_1_percent":false,"is_in_top_10_percent":false},"cited_by_percentile_year":{"min":94,"max":97},"biblio":{"volume":"34","issue":"1","first_page":"204","last_page":"211"},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T10404","display_name":"Enzyme Catalysis and Immobilization","score":0.9429000020027161,"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"}},"topics":[{"id":"https://openalex.org/T10404","display_name":"Enzyme Catalysis and Immobilization","score":0.9429000020027161,"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/T13326","display_name":"Biochemical and Structural Characterization","score":0.04430000111460686,"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/T10169","display_name":"Protein Kinase Regulation and GTPase Signaling","score":0.004399999976158142,"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"}}],"keywords":[{"id":"https://openalex.org/keywords/protein-kinase-a","display_name":"protein kinase A","score":0.9990000128746033},{"id":"https://openalex.org/keywords/heterologous-expression-in-escherichia-coli","display_name":"heterologous expression in Escherichia coli","score":0.949999988079071},{"id":"https://openalex.org/keywords/vasp","display_name":"VASP","score":0.9390000104904175},{"id":"https://openalex.org/keywords/enzyme-reuse","display_name":"enzyme reusability","score":0.843999981880188}],"concepts":[{"id":"https://openalex.org/C185592680","wikidata":"https://www.wikidata.org/wiki/Q2329","display_name":"Chemistry","level":0,"score":0.7519508600234985},{"id":"https://openalex.org/C97029542","wikidata":"https://www.wikidata.org/wiki/Q281417","display_name":"Protein kinase A","level":3,"score":0.6654330492019653},{"id":"https://openalex.org/C150109051","wikidata":"https://www.wikidata.org/wiki/Q3630494","display_name":"Autophosphorylation","level":4,"score":0.649640679359436},{"id":"https://openalex.org/C2780115692","wikidata":"https://www.wikidata.org/wiki/Q5747302","display_name":"Heterologous","level":3,"score":0.6100467443466187},{"id":"https://openalex.org/C184235292","wikidata":"https://www.wikidata.org/wiki/Q421851","display_name":"Kinase","level":2,"score":0.5892314910888672},{"id":"https://openalex.org/C11960822","wikidata":"https://www.wikidata.org/wiki/Q242736","display_name":"Phosphorylation","level":2,"score":0.5855275392532349},{"id":"https://openalex.org/C55493867","wikidata":"https://www.wikidata.org/wiki/Q7094","display_name":"Biochemistry","level":1,"score":0.5678324699401855},{"id":"https://openalex.org/C123894998","wikidata":"https://www.wikidata.org/wiki/Q425158","display_name":"Fusion protein","level":4,"score":0.5445048213005066},{"id":"https://openalex.org/C2777061661","wikidata":"https://www.wikidata.org/wiki/Q423934","display_name":"Phosphoprotein","level":3,"score":0.520232081413269},{"id":"https://openalex.org/C147816474","wikidata":"https://www.wikidata.org/wiki/Q169525","display_name":"Protein engineering","level":3,"score":0.4341529607772827},{"id":"https://openalex.org/C108305142","wikidata":"https://www.wikidata.org/wiki/Q3062286","display_name":"Heterologous expression","level":4,"score":0.43062078952789307},{"id":"https://openalex.org/C181199279","wikidata":"https://www.wikidata.org/wiki/Q8047","display_name":"Enzyme","level":2,"score":0.429006963968277},{"id":"https://openalex.org/C2780430158","wikidata":"https://www.wikidata.org/wiki/Q7685902","display_name":"Target protein","level":3,"score":0.41550832986831665},{"id":"https://openalex.org/C40767141","wikidata":"https://www.wikidata.org/wiki/Q285697","display_name":"Recombinant DNA","level":3,"score":0.40611976385116577},{"id":"https://openalex.org/C95444343","wikidata":"https://www.wikidata.org/wiki/Q7141","display_name":"Cell biology","level":1,"score":0.33878880739212036},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.13732853531837463},{"id":"https://openalex.org/C104317684","wikidata":"https://www.wikidata.org/wiki/Q7187","display_name":"Gene","level":2,"score":0.0}],"mesh":[{"descriptor_ui":"D004926","descriptor_name":"Escherichia coli","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":false},{"descriptor_ui":"D004926","descriptor_name":"Escherichia coli","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":false},{"descriptor_ui":"D010766","descriptor_name":"Phosphorylation","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D011494","descriptor_name":"Protein Kinases","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":true},{"descriptor_ui":"D011993","descriptor_name":"Recombinant Fusion Proteins","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":false},{"descriptor_ui":"D017868","descriptor_name":"Cyclic AMP-Dependent Protein Kinases","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":true},{"descriptor_ui":"D017868","descriptor_name":"Cyclic AMP-Dependent Protein Kinases","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":true}],"locations_count":3,"locations":[{"id":"doi:10.1021/acs.bioconjchem.2c00485","doi":"https://doi.org/10.1021/acs.bioconjchem.2c00485","is_oa":false,"landing_page_url":"https://doi.org/10.1021/acs.bioconjchem.2c00485","pdf_url":null,"source":{"id":"https://openalex.org/S155115410","display_name":"Bioconjugate Chemistry","issn_l":"1043-1802","issn":["1043-1802","1520-4812"],"is_oa":false,"is_in_doaj":false,"is_core":true,"listed_in":["cwts-core","dongbi-b","doyens","jufo-1","ki-jl-1","medline","norway-1","poland-100","russia-white-list-1"],"host_organization":"https://openalex.org/P4310320006","host_organization_name":"American Chemical Society","host_organization_lineage":["https://openalex.org/P4310320006"],"host_organization_lineage_names":["American Chemical Society"],"type":"journal"},"license":null,"license_id":null,"version":"publishedVersion","is_accepted":true,"is_published":true,"raw_source_name":"Bioconjugate Chemistry","raw_type":"journal-article"},{"id":"doi:10.26434/chemrxiv-2022-c6wf5","doi":"https://doi.org/10.26434/chemrxiv-2022-c6wf5","is_oa":true,"landing_page_url":"https://doi.org/10.26434/chemrxiv-2022-c6wf5","pdf_url":"https://chemrxiv.org/engage/api-gateway/chemrxiv/assets/orp/resource/item/624326eaed3780eb0d469f08/original/design-and-application-of-an-immobilized-protein-kinase.pdf","source":{"id":"https://openalex.org/S4393918830","display_name":"ChemRxiv","issn_l":"2573-2293","issn":["2573-2293"],"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","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"},{"id":"pmid:36379001","doi":null,"is_oa":false,"landing_page_url":"https://pubmed.ncbi.nlm.nih.gov/36379001","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":"Bioconjugate chemistry","raw_type":"Journal Article"}],"best_oa_location":{"id":"doi:10.26434/chemrxiv-2022-c6wf5","doi":"https://doi.org/10.26434/chemrxiv-2022-c6wf5","is_oa":true,"landing_page_url":"https://doi.org/10.26434/chemrxiv-2022-c6wf5","pdf_url":"https://chemrxiv.org/engage/api-gateway/chemrxiv/assets/orp/resource/item/624326eaed3780eb0d469f08/original/design-and-application-of-an-immobilized-protein-kinase.pdf","source":{"id":"https://openalex.org/S4393918830","display_name":"ChemRxiv","issn_l":"2573-2293","issn":["2573-2293"],"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","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"},"sustainable_development_goals":[],"sustainable_development_goals_aurora":[],"study_designs":[],"awards":[{"id":"https://openalex.org/G3487407322","display_name":null,"funder_award_id":"DNI 57850","funder_id":"https://openalex.org/F4320332454","funder_display_name":"American Chemical Society Petroleum Research Fund"},{"id":"https://openalex.org/G8114863068","display_name":"REU Site: Experimental and Computational Research in Chemistry and Biochemistry","funder_award_id":"1851844","funder_id":"https://openalex.org/F4320306076","funder_display_name":"National Science Foundation"}],"funders":[{"id":"https://openalex.org/F4320306076","display_name":"National Science Foundation","ror":"https://ror.org/021nxhr62"},{"id":"https://openalex.org/F4320332454","display_name":"American Chemical Society Petroleum Research Fund","ror":"https://ror.org/059dqb057"}],"has_content":{"pdf":true,"grobid_xml":true},"content_urls":{"pdf":"https://content.openalex.org/works/W4309166196.pdf","grobid_xml":"https://content.openalex.org/works/W4309166196.grobid-xml"},"referenced_works_count":40,"referenced_works":["https://openalex.org/W1019142424","https://openalex.org/W1521648426","https://openalex.org/W1526841021","https://openalex.org/W1682566403","https://openalex.org/W1803226853","https://openalex.org/W1970409337","https://openalex.org/W1976540841","https://openalex.org/W1982718958","https://openalex.org/W1999085006","https://openalex.org/W1999085564","https://openalex.org/W2008030719","https://openalex.org/W2015869903","https://openalex.org/W2034132443","https://openalex.org/W2038197401","https://openalex.org/W2057127856","https://openalex.org/W2057447099","https://openalex.org/W2073967813","https://openalex.org/W2077956201","https://openalex.org/W2119328689","https://openalex.org/W2131640038","https://openalex.org/W2134178604","https://openalex.org/W2138820253","https://openalex.org/W2167206773","https://openalex.org/W2167854729","https://openalex.org/W2253318299","https://openalex.org/W2397093333","https://openalex.org/W2474754435","https://openalex.org/W2560839357","https://openalex.org/W2766215052","https://openalex.org/W2773063136","https://openalex.org/W2790741808","https://openalex.org/W2890909648","https://openalex.org/W2892265724","https://openalex.org/W2906905739","https://openalex.org/W2924856415","https://openalex.org/W2954472648","https://openalex.org/W2972775142","https://openalex.org/W2973851293","https://openalex.org/W3038138578","https://openalex.org/W3125105082"],"related_works":["https://openalex.org/W6999714328","https://openalex.org/W1930219349","https://openalex.org/W1490077964","https://openalex.org/W7162760030","https://openalex.org/W1992524715","https://openalex.org/W2015237182","https://openalex.org/W2054537781","https://openalex.org/W1975830155","https://openalex.org/W2068395639","https://openalex.org/W2803046136"],"abstract_inverted_index":{"Protein":[0],"Kinase":[1],"A":[2],"(PKA)":[3],"is":[4,100],"a":[5,30,78,133,141,157,161],"biologically":[6],"important":[7],"enzyme":[8,68],"for":[9,33,102,144],"cell":[10,134],"regulation,":[11],"often":[12,64],"referred":[13],"to":[14],"as":[15,126,140],"the":[16,56,75,85,119,145],"\u201ccentral":[17],"kinase\u201d.":[18],"An":[19],"immobilized":[20,67,116,150],"PKA":[21,80,123,130],"that":[22,62,83,92],"retains":[23],"substrate":[24],"specificity":[25],"and":[26,60,128,147],"activity":[27],"would":[28],"be":[29],"useful":[31],"tool":[32],"laboratory":[34],"scientists,":[35],"enabling":[36],"targeted":[37],"phosphorylation":[38,120],"without":[39],"interference":[40],"from":[41],"downstream":[42],"kinase":[43,46],"contamination":[44],"or":[45],"autophosphorylation":[47],"in":[48,106,132],"sensitive":[49],"assays.":[50],"Moreover,":[51],"it":[52],"might":[53],"also":[54],"offer":[55],"benefits":[57],"of":[58,77,114,121,149,156],"robustness":[59],"reusability":[61],"are":[63],"associated":[65],"with":[66],"preparations.":[69],"In":[70],"this":[71],"work,":[72],"we":[73,110],"describe":[74],"creation":[76],"recombinant":[79,122],"fusion":[81,94,158],"protein":[82,93,151,159],"incorporates":[84],"HaloTag":[86],"covalent":[87],"immobilization":[88],"system.":[89],"We":[90],"demonstrate":[91,111],"design,":[95],"including":[96,118],"affinity":[97],"tag":[98],"placement,":[99],"critical":[101],"optimal":[103],"heterologous":[104],"expression":[105],"E.":[107],"coli.":[108],"Furthermore,":[109],"various":[112],"applications":[113],"our":[115],"PKA,":[117],"substrates":[124,131],"such":[125],"VASP":[127],"endogenous":[129],"lysate.":[135],"These":[136],"results":[137],"hold":[138],"promise":[139],"generalizable":[142],"strategy":[143],"production":[146],"application":[148],"kinases":[152],"involving":[153],"specific":[154],"sequestration":[155],"by":[160],"solid":[162],"support":[163],"material.":[164]},"counts_by_year":[{"year":2026,"cited_by_count":2},{"year":2024,"cited_by_count":2}],"updated_date":"2026-10-31T12:09:11.096373","created_date":"2022-11-24T00:00:00"}
