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Pechalrieu"],"technology_type":["Mass Spectrometry","diaPASEF","Bottom-up proteomics","Data-independent acquisition"],"disease":["Malignant Neoplasm Of Ovary"],"software":[""],"submitter_keywords":["Cancer biology","Natural products","Target identification","Proteomics","Ovarian cancer"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD070302"],"sample_protocol":["Cells were treated with wheldone (1 µM) or DMSO for 24 hours, washed with PBS, scrapped, and centrifuged. Cell pellets were resuspended in PBS, proteins were extracted via sonication, and protein quantification was performed via the Bradford assay (Bio-Rad). LC-MS/MS sample (20 µg in 15 µL) preparation was performed as described previously . Peptides were resuspended in water with 0.1 % formic acid (FA) and analyzed using nanoElute 2 coupled to a TimsTOF HT mass spectrometer (Bruker Daltonics). The chromatography column consisted of a 25 cm long, 150 μm i.d. microcapillary packed with 1.5 μm C18 particles (Bruker Daltonics/Pepsep) and was capped by a 20 μm emitter (Bruker Daltonics). LC solvents were 0.1 % FA in H2O (Buffer A) and 0.1 % FA in MeCN (Buffer B). Peptides were eluted into the MS at a flow rate of 500 nL/min. over a 60 min. linear-gradient (5-35 % Buffer B) at 50 °C. Data was acquired via dia-PASEF using 20 MS/MS windows (optimized using py_DiAID ) per one MS1 scan. The method covered m/z 400-1200 and scan range between 0.6 to 1.4 1/K0. Ramp time was set to 100 ms and total cycle lasted 2.23 s. Collision energy was set as a linear increase from 20 eV at 1/K0 = 0.6 to 59 eV at 1/K0 = 1.6."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["The MS data was analyzed with DIA-NN (V1.8.1), a generated human peptide library, and the Uniprot human proteome with contaminants as database. The precursor FDR was set to 1% and MBR was checked."],"omics_type":["Proteomics"],"labhead":["Alexander Adibekian"],"instrument_platform":[""],"labhead_affiliation":["Department of Chemistry, University of Illinois Chicago, Chicago, USA"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"publication":["41881190 Khin M, Saldana AC, Rangel-Grimaldo M, Raja HA, Abegg D, Ekiert J, Liu C, Misra S, Ratia K, Adibekian A, Gao Y, Kulp SK, Coss CC, Oberlies NH, Burdette JE. Natural Product Target Identification of Wheldone, a Fungal Metabolite, as a KIF11 Inhibitor in Ovarian Cancer Using the DiffPOP (Differential Protein Precipitation) Method. Mol Cell Proteomics. 2026 25(5):101558 10.1016/j.mcpro.2026.101558"],"submitter_mail":["dpechalr@uic.edu"],"submitter_affiliation":["University of Illinois Chicago"],"submitter_country":["United States"],"pubmed_abstract":["Wheldone, a fungal metabolite, was identified as a cytotoxic compound in high-grade serous ovarian cancer (HGSOC). Wheldone induced caspase 3/7-dependent apoptosis and reduced migration, invasion, and spheroid growth. Wheldone stimulated apoptosis in chemoresistant HGSOC models. Wheldone treatment caused significant downregulation of HNRNPD, a DNA repair protein, and increased DNA damage that could be blocked by N-acetyl-L-cysteine. In vivo, wheldone displayed minimal toxicity but was rapidly cleared from circulation, despite in vitro metabolic stability. Wheldone treatment in vivo did not demonstrate significant reduction in tumor burden. Therefore, in order to overcome these liabilities, it was necessary to find the protein target of wheldone so that modifications can be made to improve the drug-like characteristics of the compound. Using the drug-target interaction proteomics method, differential precipitation of proteins, wheldone was found to act as an inhibitor of Kinesin superfamily protein 11 (KIF11), a motor protein essential for mitotic spindle formation. An ATPase biochemical cell-free assay confirmed direct binding and functional inhibition of KIF11. Wheldone resulted in G2/M arrest and downstream regulation of mitotic proteins such as TPX2, AURKA, and phospho-histone H3. Proteomics after treatment of wheldone in four different HGSOC cancer cell lines all supported changes consistent with mitotic spindle assembly disruption. Further, KIF11 was one of only 13 proteins upregulated in all 4 cell lines treated. Overall, wheldone was found to be a fungal metabolite that inhibits KIF11 in chemoresistant ovarian cancer, with future studies needed to improve its pharmacokinetics and delivery."],"pubmed_title":["Natural Product Target Identification of Wheldone, a Fungal Metabolite, as a KIF11 Inhibitor in Ovarian Cancer Using the DiffPOP (Differential Protein Precipitation) Method."],"pubmed_authors":["Khin Manead M, Saldana Alejandra Cavazos AC, Rangel-Grimaldo Manuel M, Raja Huzefa A HA, Abegg Daniel D, Ekiert Julia J, Liu Chang C, Misra Sweta S, Ratia Kiira K, Adibekian Alexander A, Gao Yu Y, Kulp Samuel K SK, Coss Christopher C CC, Oberlies Nicholas H NH, Burdette Joanna E JE"],"additional_accession":[]},"is_claimable":false,"name":"Natural product target identification of wheldone as a KIF11 inhibitor in ovarian cancer using the DiffPOP (Differential Protein Precipitation) method","description":"Wheldone, a fungal metabolite, was identified as a cytotoxic compound in high-grade serous ovarian cancer (HGSOC). Wheldone triggered caspase 3/7–dependent apoptosis and reduced migration, invasion, and spheroid growth. Wheldone was also capable of triggering apoptosis in chemoresistant HGSOC models. Wheldone caused significant downregulation of HNRNPD expression, a DNA repair protein, and increased DNA damage that could be blocked by N-acetyl-L-cysteine. In vivo, wheldone displayed minimal toxicity but was rapidly cleared from circulation, despite in vitro metabolic stability. Wheldone treatment in vivo did not demonstrate significant reduction in tumor burden. Therefore, in order to overcome these liabilities, it was necessary to find the protein target of wheldone so that modifications can be made to improve the drug-like characteristics of the compound. Using the drug-target interaction proteomics method, DiffPOP (differential precipitation of proteins), wheldone was found to act as an inhibitor of KIF11, a motor protein essential for mitotic spindle formation. An ATPase biochemical cell-free assay confirmed direct binding and functional inhibition of KIF11. Wheldone resulted in G2/M arrest and downstream regulation of mitotic proteins such as TPX2, AURKA, and phospho-histone H3. Proteomics analysis after treatment of wheldone in four different HGSOC cancer cell lines all supported changes consistent with mitotic spindle assembly disruption. Further, KIF11 was one of only 13 proteins upregulated in all four cell lines treated. Overall, wheldone represents a promising lead compound for targeting KIF11 in chemoresistant ovarian cancer, with future studies needed to improve its pharmacokinetics and delivery.","dates":{"publication":"2026-05-25","submission":"2025-11-04"},"accession":"PXD070302","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:7719","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:2065263","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:10804","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","N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