{"id":"https://openalex.org/W1968525846","doi":"https://doi.org/10.1021/bi9924718","title":"Analysis of Affinity and Specificity in an EF-Hand Site Using Double Mutant Cycles","display_name":"Analysis of Affinity and Specificity in an EF-Hand Site Using Double Mutant Cycles","publication_year":2000,"publication_date":"2000-03-11","ids":{"openalex":"https://openalex.org/W1968525846","doi":"https://doi.org/10.1021/bi9924718","mag":"1968525846","pmid":"https://pubmed.ncbi.nlm.nih.gov/10736159"},"language":"en","primary_location":{"id":"doi:10.1021/bi9924718","doi":"https://doi.org/10.1021/bi9924718","is_oa":false,"landing_page_url":"https://doi.org/10.1021/bi9924718","pdf_url":null,"source":{"id":"https://openalex.org/S104038378","display_name":"Biochemistry","issn_l":"0006-2960","issn":["0006-2960","1520-4995","1943-295X"],"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-2"],"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":"Biochemistry","raw_type":"journal-article"},"volume":null,"type":"article","is_traditional":true,"indexed_in":["crossref","pubmed"],"open_access":{"is_oa":false,"oa_status":"closed","oa_url":null,"any_repository_has_fulltext":false},"authorships":[{"author_position":"first","author":{"id":"https://openalex.org/A5033089415","display_name":"Tharin M. A. Blumenschein","orcid":"https://orcid.org/0000-0002-4932-5178","observed_orcids":["https://orcid.org/0000-0002-4932-5178"]},"institutions":[{"id":"https://openalex.org/I17974374","display_name":"Universidade de S\u00e3o Paulo","ror":"https://ror.org/036rp1748","country_code":"BR","type":"education","lineage":["https://openalex.org/I17974374"]}],"countries":["BR"],"is_corresponding":false,"raw_author_name":"Tharin M. A. Blumenschein","raw_affiliation_strings":["Depto. de Bioqu\u00edmica, Instituto de Qu\u00edmica, Universidade de S\u00e3o Paulo, Av. Prof. Lineu Prestes, 748-S\u00e3o Paulo, SP-CEP 05508-900, Brazil"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Depto. de Bioqu\u00edmica, Instituto de Qu\u00edmica, Universidade de S\u00e3o Paulo, Av. Prof. Lineu Prestes, 748-S\u00e3o Paulo, SP-CEP 05508-900, Brazil","institution_ids":["https://openalex.org/I17974374"]}]},{"author_position":"last","author":{"id":"https://openalex.org/A5061301702","display_name":"Fernando C. Reinach","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I17974374","display_name":"Universidade de S\u00e3o Paulo","ror":"https://ror.org/036rp1748","country_code":"BR","type":"education","lineage":["https://openalex.org/I17974374"]}],"countries":["BR"],"is_corresponding":false,"raw_author_name":"Fernando C. Reinach","raw_affiliation_strings":["Depto. de Bioqu\u00edmica, Instituto de Qu\u00edmica, Universidade de S\u00e3o Paulo, Av. Prof. Lineu Prestes, 748-S\u00e3o Paulo, SP-CEP 05508-900, Brazil"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Depto. de Bioqu\u00edmica, Instituto de Qu\u00edmica, Universidade de S\u00e3o Paulo, Av. Prof. Lineu Prestes, 748-S\u00e3o Paulo, SP-CEP 05508-900, Brazil","institution_ids":["https://openalex.org/I17974374"]}]}],"institutions":[],"countries_distinct_count":1,"institutions_distinct_count":1,"corresponding_author_ids":[],"corresponding_institution_ids":["https://openalex.org/I17974374"],"apc_list":{"value":4500,"currency":"USD","value_usd":4500},"apc_paid":null,"fwci":0.2986,"has_fulltext":false,"cited_by_count":14,"citation_normalized_percentile":{"value":0.47386186,"is_in_top_1_percent":false,"is_in_top_10_percent":false},"cited_by_percentile_year":{"min":89,"max":94},"biblio":{"volume":"39","issue":"13","first_page":"3603","last_page":"3610"},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T10044","display_name":"Protein Structure and Dynamics","score":0.8529999852180481,"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/T10044","display_name":"Protein Structure and Dynamics","score":0.8529999852180481,"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/T13539","display_name":"thermodynamics and calorimetric analyses","score":0.023399999365210533,"subfield":{"id":"https://openalex.org/subfields/1606","display_name":"Physical and Theoretical Chemistry"},"field":{"id":"https://openalex.org/fields/16","display_name":"Chemistry"},"domain":{"id":"https://openalex.org/domains/3","display_name":"Physical Sciences"}},{"id":"https://openalex.org/T12094","display_name":"Hemoglobin structure and function","score":0.022700000554323196,"subfield":{"id":"https://openalex.org/subfields/1303","display_name":"Biochemistry"},"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/ef-hand","display_name":"EF-hand","score":0.8579999804496765},{"id":"https://openalex.org/keywords/calcium-binding","display_name":"calcium binding","score":0.7149999737739563},{"id":"https://openalex.org/keywords/magnesium-binding","display_name":"magnesium binding","score":0.9580000042915344},{"id":"https://openalex.org/keywords/double-mutant-cycles","display_name":"double mutant cycles","score":0.7369999885559082},{"id":"https://openalex.org/keywords/coupling-energy","display_name":"coupling energy","score":0.6800000071525574},{"id":"https://openalex.org/keywords/binding-free-energy","display_name":"binding free energy","score":0.6019999980926514}],"concepts":[{"id":"https://openalex.org/C143065580","wikidata":"https://www.wikidata.org/wiki/Q3285695","display_name":"Mutant","level":3,"score":0.8167800307273865},{"id":"https://openalex.org/C107824862","wikidata":"https://www.wikidata.org/wiki/Q616005","display_name":"Binding site","level":2,"score":0.6217154264450073},{"id":"https://openalex.org/C2776414213","wikidata":"https://www.wikidata.org/wiki/Q183290","display_name":"Serine","level":3,"score":0.5487918257713318},{"id":"https://openalex.org/C2780283098","wikidata":"https://www.wikidata.org/wiki/Q4688960","display_name":"Affinities","level":2,"score":0.47663816809654236},{"id":"https://openalex.org/C2779856020","wikidata":"https://www.wikidata.org/wiki/Q218642","display_name":"Alanine","level":3,"score":0.47174131870269775},{"id":"https://openalex.org/C185592680","wikidata":"https://www.wikidata.org/wiki/Q2329","display_name":"Chemistry","level":0,"score":0.4682800769805908},{"id":"https://openalex.org/C501734568","wikidata":"https://www.wikidata.org/wiki/Q42918","display_name":"Mutation","level":3,"score":0.453387588262558},{"id":"https://openalex.org/C207583985","wikidata":"https://www.wikidata.org/wiki/Q128723","display_name":"Wild type","level":4,"score":0.445538729429245},{"id":"https://openalex.org/C71240020","wikidata":"https://www.wikidata.org/wiki/Q186011","display_name":"Stereochemistry","level":1,"score":0.4053891897201538},{"id":"https://openalex.org/C12554922","wikidata":"https://www.wikidata.org/wiki/Q7100","display_name":"Biophysics","level":1,"score":0.3938716948032379},{"id":"https://openalex.org/C515207424","wikidata":"https://www.wikidata.org/wiki/Q8066","display_name":"Amino acid","level":2,"score":0.3718157708644867},{"id":"https://openalex.org/C55493867","wikidata":"https://www.wikidata.org/wiki/Q7094","display_name":"Biochemistry","level":1,"score":0.3643324077129364},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.2677726745605469},{"id":"https://openalex.org/C104317684","wikidata":"https://www.wikidata.org/wiki/Q7187","display_name":"Gene","level":2,"score":0.09678667783737183},{"id":"https://openalex.org/C181199279","wikidata":"https://www.wikidata.org/wiki/Q8047","display_name":"Enzyme","level":2,"score":0.06453996896743774}],"mesh":[{"descriptor_ui":"D000595","descriptor_name":"Amino Acid Sequence","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D000818","descriptor_name":"Animals","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D001224","descriptor_name":"Aspartic Acid","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":false},{"descriptor_ui":"D001224","descriptor_name":"Aspartic Acid","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":false},{"descriptor_ui":"D001665","descriptor_name":"Binding Sites","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":false},{"descriptor_ui":"D002118","descriptor_name":"Calcium","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":false},{"descriptor_ui":"D002118","descriptor_name":"Calcium","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":false},{"descriptor_ui":"D002645","descriptor_name":"Chickens","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D004734","descriptor_name":"Energy Metabolism","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":false},{"descriptor_ui":"D008274","descriptor_name":"Magnesium","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":false},{"descriptor_ui":"D008274","descriptor_name":"Magnesium","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":false},{"descriptor_ui":"D008969","descriptor_name":"Molecular Sequence Data","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D013816","descriptor_name":"Thermodynamics","qualifier_ui":null,"qualifier_name":null,"is_major_topic":false},{"descriptor_ui":"D016297","descriptor_name":"Mutagenesis, Site-Directed","qualifier_ui":null,"qualifier_name":null,"is_major_topic":true},{"descriptor_ui":"D018698","descriptor_name":"Glutamic Acid","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":false},{"descriptor_ui":"D018698","descriptor_name":"Glutamic Acid","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":false},{"descriptor_ui":"D018994","descriptor_name":"Myosin Light Chains","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":true},{"descriptor_ui":"D018994","descriptor_name":"Myosin Light Chains","qualifier_ui":"Q000378","qualifier_name":"metabolism","is_major_topic":false},{"descriptor_ui":"D018994","descriptor_name":"Myosin Light Chains","qualifier_ui":"Q000737","qualifier_name":"chemistry","is_major_topic":true},{"descriptor_ui":"D020832","descriptor_name":"EF Hand Motifs","qualifier_ui":"Q000235","qualifier_name":"genetics","is_major_topic":true}],"locations_count":3,"locations":[{"id":"doi:10.1021/bi9924718","doi":"https://doi.org/10.1021/bi9924718","is_oa":false,"landing_page_url":"https://doi.org/10.1021/bi9924718","pdf_url":null,"source":{"id":"https://openalex.org/S104038378","display_name":"Biochemistry","issn_l":"0006-2960","issn":["0006-2960","1520-4995","1943-295X"],"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-2"],"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":"Biochemistry","raw_type":"journal-article"},{"id":"pmid:10736159","doi":null,"is_oa":false,"landing_page_url":"https://pubmed.ncbi.nlm.nih.gov/10736159","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":"Biochemistry","raw_type":"Journal Article"},{"id":"pmh:oai:ueaeprints.uea.ac.uk:21381","doi":null,"is_oa":false,"landing_page_url":"https://ueaeprints.uea.ac.uk/id/eprint/21381/","pdf_url":null,"source":{"id":"https://openalex.org/S4306400384","display_name":"UEA Digital Repository (University of East Anglia)","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":"https://openalex.org/I1118541","host_organization_name":"University of East Anglia","host_organization_lineage":["https://openalex.org/I1118541"],"host_organization_lineage_names":[],"type":"repository"},"license":null,"license_id":null,"version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":null,"raw_type":"Article"}],"best_oa_location":null,"sustainable_development_goals":[],"study_designs":[],"awards":[],"funders":[],"has_content":{"grobid_xml":false,"pdf":false},"content_urls":null,"referenced_works_count":14,"referenced_works":["https://openalex.org/W1503782530","https://openalex.org/W1571243084","https://openalex.org/W1731598977","https://openalex.org/W1974464563","https://openalex.org/W1983265552","https://openalex.org/W1983659508","https://openalex.org/W1985322317","https://openalex.org/W1991622845","https://openalex.org/W2012396129","https://openalex.org/W2019213007","https://openalex.org/W2021577269","https://openalex.org/W2030590011","https://openalex.org/W2060041029","https://openalex.org/W2097813999"],"related_works":["https://openalex.org/W7120773503","https://openalex.org/W1551583508","https://openalex.org/W1524268590","https://openalex.org/W2913555249","https://openalex.org/W2007776478","https://openalex.org/W98746484","https://openalex.org/W4384831369","https://openalex.org/W1974019900","https://openalex.org/W2091460691","https://openalex.org/W2912750279"],"abstract_inverted_index":{"The":[0,73],"effects":[1],"of":[2,11,34,67,112,146,169],"three":[3,69],"mutations":[4,70,107],"on":[5,138],"the":[6,35,54,68,80,90,113,122,127,139,143,147,150,157,164,207,245],"EF-hand":[7],"Ca(2+)/Mg(2+)":[8],"binding":[9,130,172,180,193,212],"site":[10],"smooth":[12],"muscle":[13],"myosin":[14],"regulatory":[15],"light":[16],"chain":[17],"(RLC)":[18],"were":[19,71],"studied:":[20],"D5S,":[21],"in":[22,31,38,47,52,56,129,156,230,234,244],"which":[23,39,53,194],"an":[24,40,196,227],"aspartate":[25,41,55,228],"is":[26,42,59,205,217,229],"replaced":[27,43,60],"by":[28,44,61],"a":[29,45,62,186],"serine":[30],"position":[32,48,57,231,235],"5":[33],"loop;":[36],"D9E,":[37],"glutamate":[46],"9;":[49],"and":[50,77,79,97,183,215],"D12E,":[51],"12":[58],"glutamate.":[63],"All":[64,89,111],"possible":[65],"combinations":[66],"produced.":[72],"single":[74],"mutants":[75,91,114,170],"D5S":[76,108,182,214],"D9E":[78,184,216],"double":[81,246],"mutant":[82,135,247],"D5S/D9E":[83],"have":[84,98,116,176,185],"low":[85],"affinity":[86,118],"for":[87,119,166,191,210],"Ca(2+).":[88],"containing":[92,106],"mutation":[93,203],"D12E":[94,204],"are":[95,153,162],"Ca(2+)-specific":[96],"higher":[99],"affinities":[100],"than":[101,121],"wild":[102],"type,":[103],"even":[104],"when":[105,171,179],"or":[109],"D9E.":[110],"studied":[115],"lower":[117],"Mg(2+)":[120,192,211,242],"wild-type":[123],"protein.":[124],"As":[125],"expected,":[126],"changes":[128],"free":[131],"energy":[132,190,209],"that":[133,221],"each":[134],"produces":[136],"depend":[137],"residues":[140],"present":[141],"at":[142],"other":[144],"positions":[145,152],"site,":[148],"since":[149],"mutated":[151],"very":[154],"close":[155],"protein":[158],"structure.":[159],"Coupling":[160],"energies":[161],"about":[163],"same":[165],"all":[167],"pairs":[168],"Ca(2+),":[173],"but":[174],"can":[175],"different":[177],"values":[178],"Mg(2+).":[181],"large":[187],"negative":[188],"coupling":[189,208],"suggests":[195],"interaction":[197,223],"between":[198,213],"these":[199],"two":[200],"positions.":[201],"When":[202],"present,":[206],"much":[218],"lower,":[219],"suggesting":[220],"this":[222],"occurs":[224],"only":[225],"if":[226],"12.":[232],"Glutamate":[233],"9":[236],"may":[237],"be":[238],"able":[239],"to":[240],"coordinate":[241],"directly":[243],"D5S/D9E.":[248]},"counts_by_year":[{"year":2025,"cited_by_count":1},{"year":2017,"cited_by_count":1},{"year":2015,"cited_by_count":1}],"updated_date":"2026-11-04T10:54:05.112534","created_date":"2016-06-24T00:00:00"}
