{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:46:41Z","timestamp":1785851201252,"version":"3.56.0"},"reference-count":79,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100006492","name":"Division of Intramural Research, National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI173295"],"award-info":[{"award-number":["R01AI173295"]}],"id":[{"id":"10.13039\/100006492","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2026,8,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      In the malaria parasite\n                      <jats:italic toggle=\"yes\">Plasmodium falciparum<\/jats:italic>\n                      , the essential chaperone\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 regulates proteostasis through protein folding, but its domain-specific functions remain poorly defined. The protein contains an N-terminal nucleotide-binding domain (NBD) and a C-terminal substrate-binding domain (SBD), where selective inhibitors are needed to probe domain-specific functions and advance\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 as an antimalarial drug target. Here, we identified small molecules that bind\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 using a high-throughput thermal shift screen, an unbiased approach that enables the discovery of ligands targeting any\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 domain. Molecules were prioritized by their affinity to\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 and anti-\n                      <jats:italic toggle=\"yes\">Plasmodium<\/jats:italic>\n                      activity. These efforts led to the characterization of AMK3 and AMK4, which bind to\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 with low-micromolar affinity and exhibit selectivity (&gt;25-fold) for\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 over the human homolog HSPA1A based on microscale thermophoresis. The\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 binding regions of AMK3 and AMK4 were isolated using a combination of proteomic and biochemical methods, which demonstrated that AMK3 binds to the NBD and competes with ATP binding. In contrast, AMK4 binds to the SBD and leads to disruption in peptide binding. A molecular dynamic-facilitated structure-activity relationship (SAR) screen was performed on AMK3, yielding a compound with retained anti-\n                      <jats:italic toggle=\"yes\">Plasmodium<\/jats:italic>\n                      activity but reduced host cytotoxicity. This study identifies species-selective N-terminal and C-terminal inhibitors of\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1, and suggests druggable binding pockets on the C-terminal domain that may be exploited for the disruption of essential protein-protein interactions. These domain-specific inhibitors are useful starting points for the development of probes to advance our knowledge of\n                      <jats:italic toggle=\"yes\">Pf<\/jats:italic>\n                      Hsp70-1 functions during infection and as therapeutics to treat malaria.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.00466-26","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T18:29:28Z","timestamp":1783016968000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Development of domain-specific probes of\n                    <i>Plasmodium falciparum<\/i>\n                    heat shock protein 70-1"],"prefix":"10.1128","volume":"70","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6879-0588","authenticated-orcid":false,"given":"Aaron M.","family":"Keeler","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-1694-7277","authenticated-orcid":false,"given":"Gaini","family":"Ibrasheva","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth G.","family":"Boger","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueqi","family":"Chen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Porter E.","family":"Petruzziello","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erin A.","family":"Schroeder","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03njmea73","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher R.","family":"Mansfield","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03njmea73","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kayla","family":"Sylvester","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03njmea73","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cameron D.","family":"Fleming","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip F.","family":"Hughes","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03njmea73","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Pharmacology and Cancer Biology, Duke University Medical Center","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy A. J.","family":"Haystead","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03njmea73","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Pharmacology and Cancer Biology, Duke University Medical Center","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael C.","family":"Fitzgerald","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6664-8844","authenticated-orcid":true,"given":"Emily R.","family":"Derbyshire","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00py81415","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Duke University","place":["Durham, USA"]},{"id":[{"id":"https:\/\/ror.org\/03njmea73","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center","place":["Durham, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.lanmic.2025.101073"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-019-0306-2"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2875-13-118"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2022.02.009"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.biocel.2007.02.011"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.161"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-019-0133-3"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.072918107"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0903503106"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-004-4464-6"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2019.05.026"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/bies.20107"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-04252-1"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1914376117"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.2174\/1568026616666160413140911"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2003.09.010"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2003.10.038"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12551-019-00563-w"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2009.01.024"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(91)90163-z"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.56773"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.3390\/cancers13122936"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2014.10.016"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/ml400204n"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms24044083"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200802801"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm801627a"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2014-0138"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1515\/BC.2011.040"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12192-017-0797-6"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.3390\/molecules22122139"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12192-021-01212-6"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1107\/S2053230X21007378"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)41940-5"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0129445"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22094417"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1118370109"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-62703-706-8_21"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0135326"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA118.002691"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03819-2"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btx350"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.21334"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv315"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.1c00203"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA119.009686"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.275057"},{"key":"e_1_3_3_49_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0137298"},{"key":"e_1_3_3_50_2","doi-asserted-by":"publisher","DOI":"10.1096\/fj.201901741R"},{"key":"e_1_3_3_51_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23031771"},{"key":"e_1_3_3_52_2","doi-asserted-by":"publisher","DOI":"10.1155\/2022\/1178228"},{"key":"e_1_3_3_53_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2123238119"},{"key":"e_1_3_3_54_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1003379"},{"key":"e_1_3_3_55_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2022.06.006"},{"key":"e_1_3_3_56_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2007.12.009"},{"key":"e_1_3_3_57_2","doi-asserted-by":"publisher","DOI":"10.1177\/1087057110380571"},{"key":"e_1_3_3_58_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2004.04.031"},{"key":"e_1_3_3_59_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0103518"},{"key":"e_1_3_3_60_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02129-25"},{"key":"e_1_3_3_61_2","doi-asserted-by":"publisher","DOI":"10.2174\/138161213804143699"},{"key":"e_1_3_3_62_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi002656a"},{"key":"e_1_3_3_63_2","doi-asserted-by":"publisher","DOI":"10.1021\/cb500236y"},{"key":"e_1_3_3_64_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2014.11.007"},{"key":"e_1_3_3_65_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.11.002"},{"key":"e_1_3_3_66_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.02.020"},{"key":"e_1_3_3_67_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-022-01488-1"},{"key":"e_1_3_3_68_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature19804"},{"key":"e_1_3_3_69_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2021.114170"},{"key":"e_1_3_3_70_2","doi-asserted-by":"publisher","DOI":"10.1021\/jasms.9b00041"},{"key":"e_1_3_3_71_2","doi-asserted-by":"publisher","DOI":"10.1021\/ct400314y"},{"key":"e_1_3_3_72_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cpc.2012.09.022"},{"key":"e_1_3_3_73_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.9b02372"},{"key":"e_1_3_3_74_2","unstructured":"CaseDA Aktulga HM Belfon KAA Ben-Shalom I Brozell DS Cerutti TE Cheatham III GA Cisneros VWD Cruzeiro TA Darden RE et al.. 2021. Amber 2021. https:\/\/ambermd.org\/doc12\/Amber21.pdf."},{"key":"e_1_3_3_75_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmgm.2005.12.005"},{"key":"e_1_3_3_76_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20035"},{"key":"e_1_3_3_77_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jctc.5b00255"},{"key":"e_1_3_3_78_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.464397"},{"key":"e_1_3_3_79_2","doi-asserted-by":"publisher","DOI":"10.1016\/0021-9991(77)90098-5"},{"key":"e_1_3_3_80_2","doi-asserted-by":"publisher","DOI":"10.1021\/ct400341p"}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aac.00466-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aac.00466-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:02:34Z","timestamp":1785848554000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/aac.00466-26"}},"subtitle":[],"editor":[{"given":"Audrey","family":"Odom John","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":79,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2026,8,5]]}},"alternative-id":["10.1128\/aac.00466-26"],"URL":"https:\/\/doi.org\/10.1128\/aac.00466-26","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"value":"0066-4804","type":"print"},{"value":"1098-6596","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]},"assertion":[{"value":"2026-03-27","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-12","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-02","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00466-26"}}