{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Tabular":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/translatome-combined_protein.tsv","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/total-gametocyte-proteome-report.pr_matrix.tsv","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/translatome-combined_peptide.tsv","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/total-gametocyte-proteome-report.pg_matrix.tsv"],"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/checksum.txt"],"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/JH-EAdS-WT-2_S3-B10_1_2993.d.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/JH-EAdS-WT-1_S3-B9_1_2992.d.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/JH-ES-C-2_S4-F2_1_1454.d.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/JH-EAdS-WT-3_S3-B11_1_2994.d.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/JH-ES-B_S4-F1_1_1451.d.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD075878/JH-ES-A_S4-E12_1_1450.d.zip"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["michael.delves@lshtm.ac.uk"],"submitter":["Jack Houghton"],"technology_type":["Data-dependent acquisition","Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition"],"disease":["Malaria"],"software":[""],"submitter_keywords":["Plasmodium falciparum","Translatome","Malaria"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD075878"],"sample_protocol":["The frozen gametocyte pellets were lysed in cold lysis buffer consisting of PBS supplemented with 1% Triton X-100, 0.01% SDS, and EDTA-free HaltTM protease inhibitor cocktail. The lysates were incubated on ice for one hour, then sonicated using an ultrasonic bath (Grant Instruments Cambridge Ltd) at full power for 30 seconds, repeated three times with 30-second intervals. Following sonication, samples were centrifuged at 16,000 × g for 30 minutes at 4°C, and the soluble fraction was collected. Protein concentration was determined using the Pierce BCA Protein Assay Kit. The soluble fractions were either directly processed for whole proteome analysis or used for click chemistry pulldown enrichment. The soluble fraction was incubated at room temperature for one hour on a shaker with a click reaction mix composed of 100 µM PEG4 carboxamide-propargyl biotin (Thermo Fisher Scientific), 1 mM copper sulfate (Scientific Laboratory Supplies), 1 mM of tris(2-carboxyethyl)phosphine hydrochloride (TCEP, Promega UK Ltd), and 100 µM tris((1-benzyl-4-triazolyl)methyl)amine (TBTA, Cambridge Bioscience). The reaction was quenched with 5 mM EDTA. Proteins were precipitated with acetonitrile and resuspended in 50 mM HEPES buffer (pH 8.0) containing 0.2% SDS, followed by incubation with NeutrAvidin agarose beads (ThermoFisher Scientific) for one hour at room temperature on a shaker. The beads were washed four times with the same buffer before proceeding to on-bead digestion. All protein samples, including whole proteome and enriched fractions, were reduced with 10 mM TCEP and alkylated with 40 mM 2-chloroacetamide (CAA), followed by overnight digestion at 37°C using sequencing-grade modified trypsin (Promega UK Ltd)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["DIA data from whole proteome samples were processed using DIA-NN (version 1.8.1)14 in library-free mode. The analysis used the Plasmodium falciparum 3D7 protein database downloaded from UniProt on 15 December 2023, which included 5374 proteins and 246 common contaminants. Deep learning-based prediction of spectra and retention times was enabled. Trypsin specificity was applied, allowing one missed cleavage, with carbamidomethylation of cysteine as a fixed modification and oxidation (M), N-terminal acetylation, and N-terminal excision set as variable modifications, with up to two variable modifications per peptide. Match-between-runs (MBR) was enabled, and quantification was performed using the \"Robust LC (high precision)\" strategy. Heuristic protein inference was disabled, and both mass and MS1 accuracy were set to 0. DDA data from the click pulldown samples were analysed using the FragPipe suite (version 5.0.0) with the built-in “LFQ MBR” (Label-Free Quantification with Match-Between-Runs) workflow. Raw .d files were processed with MSFragger (version 3.8)15, using strict trypsin digestion with up to two missed cleavages allowed, and a precursor ion tolerance of 20 ppm. Carbamidomethylation of cysteine was set as a fixed modification, while oxidation (M) and N-terminal acetylation were defined as variable modifications. Trimming of N-terminal methionine residues was enabled. The same P. falciparum 3D7 database used for DIA-NN was also used here, supplemented with 50% decoy sequences and common contaminants via the inbuilt FragPipe utility. Peptide-spectrum matches were validated using Percolator (version 3.5)16, and protein inference and FDR filtering were performed with Philosopher (version 5.0.0)17. Label-free quantification was conducted using IonQuant (version 5.0.0)18, with match-between-runs enabled and the minimum number of ions for quantification set as 1."],"omics_type":["Proteomics"],"labhead":["Michael Delves"],"instrument_platform":[""],"labhead_affiliation":["London School of Hygine and Tropical Medicine"],"submission_type":["PARTIAL"],"species":["Plasmodium Falciparum"],"submitter_mail":["j.houghton@imperial.ac.uk"],"publication":["10.64898/2026.03.24.713170"],"submitter_affiliation":["Imperial College London"],"submitter_country":["United Kingdom"],"additional_accession":[]},"is_claimable":false,"name":"The translatome of quiescent Plasmodium falciparum gametocytes reveals parasite pyridoxal 5’-phosphate (PLP) biosynthesis is essential for efficient mosquito stage development","description":"The ability of Plasmodium falciparum gametocytes to remain quiescent within the vertebrate host but poised for rapid onward development in the mosquito is an adaptation essential to maximise the onward spread of malaria. In this dormant state, mature infectious stage V gametocytes are largely unaffected by most antimalarial drugs and our limited understanding of how gametocytes prepare for mosquito transmission has hindered the identification of new molecular targets for transmission-blocking therapeutics. In this study, we move beyond the total proteome of gametocytes and define the translatome of mature stage V gametocytes using L-azidohomoalanine incorporation into nascent proteins, click chemistry purification and proteomic analysis. We identify the proteins and pathways that gametocytes sustain in preparation for transmission during this dormant period and through genetic disruption, we validate this approach by demonstrating the importance of parasite pyridoxal 5’-phosphate biosynthesis for mosquito transmission.","dates":{"publication":"2026-03-25","submission":"2026-03-19"},"accession":"PXD075878","cross_references":{"TAXONOMY":["NEWT:6945","NEWT:184922","NEWT:6703","NEWT:3555","NEWT:71647","NEWT:2","NEWT:157546","NEWT:474186","NEWT:35554","NEWT:38942","NEWT:307972","NEWT:32046","NEWT:1496","NEWT:544496","NEWT:2102","NEWT:2042546","NEWT:259447","NEWT:45351","NEWT:445974","NEWT:43179","NEWT:4513","NEWT:180454","NEWT:5722","NEWT:1247","NEWT:6938","NEWT:1129","NEWT:55153","NCBITaxon:10407","NEWT:1736309","NEWT:309800","NEWT:281395","NEWT:10360","NEWT:1211601","NEWT:876138","NEWT:47664","NEWT:317548","NEWT:3654","NEWT:237561","NEWT:5833","NEWT:6928","NEWT:10036","NEWT:36745","NEWT:498019","NEWT:1351","NEWT:1438992","NEWT:1352","NEWT:2649997","NEWT:272563","NEWT:224326","NEWT:1333499","NCBITaxon:79857","NEWT:1096976","NEWT:82688","NEWT:95648","NEWT:3885","NEWT:3888","NEWT:5821","NEWT:1589","NEWT:135622","NCBITaxon:4896","NEWT:6915","NEWT:3649","NEWT:101510","NEWT:28903","NEWT:3880","NEWT:272559","NEWT:28909","NEWT:515849","NEWT:3641","NEWT:383379","NEWT:466585","NEWT:10029","NEWT:913645","NEWT:1000589","NEWT:85963","NEWT:85962","NEWT:143361","NEWT:317447","NEWT:4688","NEWT:7955","NEWT:7959","NEWT:2261","NEWT:31156","NEWT:398580","NEWT:4442","NEWT:4565","NEWT:1264690","NEWT:515619","NEWT:192875","NEWT:34305","NEWT:59729","NEWT:2164133","NCBITaxon:183674","NEWT:1316931","NEWT:224308","NEWT:167387","NEWT:84645","NEWT:44586","NEWT:626528","NEWT:3347","NEWT:511145","NEWT:139927","NEWT:4558","NEWT:209285","NEWT:5888","NEWT:211586","NEWT:747078","NEWT:1282","NEWT:1283","NEWT:931281","NEWT:4550","NEWT:1000561","NEWT:2133","NEWT:294381","NEWT:197","NEWT:1390363","NEWT:77133","NEWT:288705","NEWT:29176","NEWT:145481","NCBITaxon:79824","NEWT:4787","NCBITaxon:4563","NEWT:5755","NEWT:44689","NEWT:3218","NEWT:5759","NEWT:1736231","NEWT:556182","NEWT:1270","NEWT:374990","NEWT:1392","NEWT:498217","NEWT:156471","NEWT:2242","NEWT:4784","NEWT:11320","NEWT:360106","NEWT:156476","NEWT:286","NEWT:391619","NEWT:360104","NEWT:246196","NEWT:287","NEWT:246197","NEWT:10117","NEWT:10239","NEWT:10116","NEWT:1280","NEWT:1735272","NEWT:83334","NEWT:83332","NEWT:44685","NEWT:317513","NEWT:161934","NEWT:1148","NEWT:580240","NEWT:5508","NEWT:294128","NEWT:5507","NEWT:11676","NEWT:55571","NEWT:35500","NEWT:1140","NEWT:100226","NEWT:4530","NEWT:1143","NEWT:4896","NEWT:75058","NEWT:13616","NEWT:1390","NEWT:1094343","NEWT:1336795","NEWT:644042","NEWT:296543","NEWT:316435","NEWT:42157","NEWT:1773","NEWT:1895","NEWT:1182590","NEWT:3712","NEWT:82380","NEWT:1034304","NEWT:105023","NEWT:866628","NEWT:935293","NEWT:64152","NEWT:4924","NEWT:749200","NEWT:375146","NEWT:9378","NEWT:990346","NEWT:145953","NEWT:257309","NEWT:100816","NEWT:263","NEWT:230741","NEWT:52283","NEWT:284812","NEWT:8175","NCBITaxon:1313","NEWT:43330","NEWT:1603293","NEWT:408169","NEWT:44544","NEWT:47946","NEWT:4911","NEWT:645463","NEWT:3702","NEWT:129249","NEWT:1077286","NEWT:243277","NEWT:990119","NEWT:7067","NEWT:2850","NEWT:1111708","NEWT:408172","NEWT:227321","NEWT:408170","NEWT:493760","NEWT:106590","NEWT:260710","NEWT:257313","NEWT:3827","NEWT:400772","NEWT:3708","NEWT:128161","NEWT:332648","NEWT:930945","NEWT:106592","NEWT:536231","NEWT:46704","NEWT:1436733","NEWT:460519","NEWT:1187947","NEWT:572307","NEWT:1432138","NEWT:269796","NEWT:10312","NEWT:1424507","NCBITaxon:1773","NEWT:1194599","NEWT:272844","NEWT:54571","NEWT:9598","NEWT:8030","NEWT:1513458","NCBITaxon:40559","NEWT:1639","NEWT:188229","NEWT:3818","NEWT:480","NEWT:4909","NEWT:67767","NEWT:759272","NEWT:432359","NEWT:46835","NEWT:1182263","NEWT:109757","NEWT:943146","NEWT:2711","NEWT:300852","NEWT:1502","NEWT:376686","NEWT:95486","NEWT:9103","NEWT:508771","NEWT:1883446","NEWT:29159","NEWT:253","NEWT:10306","NCBITaxon:2759","NEWT:1233435","NEWT:93061","NEWT:8022","NEWT:145943","NCBITaxon:4932","NEWT:595536","NEWT:240906","NEWT:593117","NEWT:89920","NEWT:29158","NEWT:3635","NEWT:5811","NEWT:235443","NEWT:180923","NEWT:108458","NEWT:272623","NEWT:272624","NEWT:411483","NEWT:884019","NEWT:198215","NEWT:411490","NEWT:983964","NEWT:118499","NEWT:169963","NEWT:32644","NEWT:527796","NEWT:225117","NEWT:746128","NEWT:499175","NEWT:109779","NEWT:43665","NEWT:1266464","NEWT:1715989","NEWT:476272","NEWT:3747","NEWT:195051","NEWT:367830","NEWT:1255228","NEWT:178616","NEWT:649908","NEWT:410289","NEWT:373153","NEWT:375451","NEWT:352472","NEWT:357","NEWT:1071661","NEWT:360094","NEWT:470","NEWT:41364","NEWT:1313","NEWT:411469","NEWT:84023","NEWT:1679","NEWT:559292","NEWT:39491","NCBITaxon:5811","NEWT:29491","NEWT:411464","NEWT:411460","NEWT:2014887","NEWT:2762","NEWT:1174673","NEWT:1328426","NEWT:562","NEWT:411470","NEWT:33952","NEWT:2094720","NCBITaxon:2697049","NEWT:571256","NCBITaxon:138","NEWT:40483","NEWT:28038","NEWT:1663","NEWT:1423","NEWT:4932","NEWT:3603","NEWT:2759","NEWT:3847","NEWT:38293","NEWT:1428","NEWT:327159","NEWT:178876","NEWT:1660","NEWT:327160","NEWT:573","NEWT:9031","NEWT:1872122","NEWT:7091","NEWT:108931","NE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