Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Total_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Digested_04.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Digested_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Total_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Digested_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Digested_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Digested_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Digested_04.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Digested_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Digested_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Total_04.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Total_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Total_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Total_04.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Healthy_Total_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/2022-07-09_MS_MG_Albino_Total_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD051372/combined.zipprimaryOK200marta.giacomello@unipd.itFrancescopaolo IavaroneMass SpectrometryTop-down proteomicsMitochondriaMelanosomeMffhttps://www.ebi.ac.uk/pride/archive/projects/PXD051372Nerve CordCell CultureAll the experiments were performed in a labeling free setting. Proteins were precipitate in acetone and then reduced and alkylated in a solution of 6M Guanidine-HCl, 5mM TCEP, and 20mM chloroacetamide. Peptides were obtained digesting proteins with LysC (WAKO) for 3hrs at 37°C and with the endopeptidase sequencing-grade Trypsin (Promega) overnight at 37°C. Collected peptide mixtures were concentrated and desalted using the Stop and Go Extraction (STAGE) technique. Instruments for LC MS/MS analysis consisted of a NanoLC 1200 coupled via a nano-electrospray ionization source to the quadrupole-based Q Exactive HF benchtop mass spectrometer. Peptide separation was carried out according to their hydrophobicity on a home-made chromatographic column, 75m ID, 8Um tip, 350mm bed packed with Reprosil-PUR, C18-AQ, 1.9m particle size, 120 Angstrom pore size, using a binary buffer system consisting of solution A: 0.1% formic acid and B: 80% acetonitrile, 0.1% formic acid.PrideNot availableAll acquired raw files were processed using MaxQuant (1.6.2.10) and the implemented Andromeda search engine. For protein assignment, spectra were correlated with the mus musculus (v. 2021) including a list of common contaminants. Searches were performed with tryptic specifications and default settings for mass tolerances for MS and MS/MS spectra. The other parameters were set as follows: Fixed modifications: Carbamidomethyl (C); Variable modifications: Oxidation, Acetyl (N-term); Digestion: Trypsin; Minimum peptide length = 7; Maximal peptide mass = 470 Da. The false discovery rate for proteins and peptide-spectrum = 1%. For further analysis, the Perseus software (1.6.2.3) was used and first filtered for contaminants and reverse entries as well as proteins that were only identified by a modified peptide [First filter]. The LFQ Ratios were set in logarithmic scale, grouped, and filtered for the minimum valid number (min. 3 in at least one group) [Second filter]. Missing values have been replaced by random numbers that are drawn from a normal distribution. Two-sample t-test analysis was performed using FDR = 0.05. Proteins with Log2 Difference≥ ± 0.5 and q-value <0.05 were considered significantly enrichedProteomicsMarta GiacomelloUniversity of PadovaPARTIALMus Musculus (mouse)f.iavarone@tigem.it41832149 Magalhães Rebelo AP, Maracani A, Greco S, Dal Bello F, Santorelli L, Gerdol M, Pallavicini A, Knedlik T, Schiavon S, Scorrano L, Goff PS, Frezza C, Sviderskaya EV, Grumati P, Giacomello M. MFF budding from mitochondria regulates melanosome size and maturation. Nat Commun. 2026 17(1):3932 10.1038/s41467-026-70572-3TIGEMItalyMelanosomes are lysosome-related organelles that produce and accumulate melanin. Their maturation is regulated through interactions with mitochondria and involves the export and recycling of proteins via tubular transport and fission events whose mechanisms are unknown. Here, we demonstrate that the mitochondrial fission factor protein (MFF) is involved in melanosome fission. MFF is trafficked between mitochondria and melanosomes and locates at melanosome fission events. Upon downregulation of MFF, but not of dynamin-related protein 1 (DRP1), melanosomes enlarge, intracellular melanin accumulates, and melanosomal lumenal catabolism increases, indicating that MFF-dependent melanosome fission is required for their maturation. We show that MFF interacts with regulators of the ARP2/3 complex, which drives F-actin nucleation. Actin filaments accumulate between melanosomes at MFF-enriched membrane constriction sites, and silencing of ARP2/3 subunits mimics the increase in melanosome size. MFF regulates actin-dependent fission of melanosomes via the ARP2/3 complex, indicating an extramitochondrial function for MFF in the regulation of melanosome homeostasis.MFF budding from mitochondria regulates melanosome size and maturation.Magalhães Rebelo Ana Paula AP, Maracani Aurora A, Greco Samuele S, Dal Bello Federica F, Santorelli Lucia L, Gerdol Marco M, Pallavicini Alberto A, Knedlik Tomas T, Schiavon Sara S, Scorrano Luca L, Goff Philip S PS, Frezza Christian C, Sviderskaya Elena V EV, Grumati Paolo P, Giacomello Marta MfalseDelivery of MFF through mitochondria-derived vesicles regulates melanosome maturationMelanosomes are lysosome-associated organelles that produce and accumulate melanin. Their biogenesis begins with the trafficking of melanogenic proteins from the endo-lysosomal system into non-pigmented vacuoles derived from endosomes. Subsequently, melanosome maturation proceeds with the export and recycling of non-required cargo, through tubular carriers and vesicular fission derived from melanosome membranes. Although constriction events have been observed at melanosome membranes, the mechanisms underlying their fission remain unclear. We found that MFF is delivered to melanosomes by mitochondria-derived vesicles (MDVs) and that it is located at melanosome membrane fission sites. Downregulation of MFF leads to an increase in the size of early melanosomes, accumulation of intracellular melanin, and increased melanosome lumen catabolism. In melanocytes, MFF interactome highlighted its interplay with a complex network of cytoskeletal factors. Inhibition of actin nucleation is indeed sufficient to restore the effects on melanosome size. Our data show for the first time that MDVs-mediated MFF trafficking to melanosomes is necessary for their membrane remodeling and fission, promoting the transition from early to mature stage melanosomes.2026-05-252024-04-11PXD051372NEWT:6945NEWT:3555NEWT:2NEWT:157546NEWT:35554NEWT:38942NEWT:307972NEWT:32046NEWT:544496NEWT:2042546NEWT:45351NEWT:43179NEWT:4513NEWT:5722NEWT:55153NCBITaxon:10407NEWT:1736309NEWT:309800NEWT:1211601NEWT:876138NEWT:237561NEWT:5833NEWT:6928NEWT:10036NEWT:36745NEWT:1351NEWT:1438992NEWT:2649997NEWT:272563NEWT:224326NCBITaxon:79857NEWT:1096976NEWT:95648NEWT:3885NEWT:3888NEWT:1589NEWT:135622NCBITaxon:4896NEWT:6915NEWT:3649NEWT:101510NEWT:3880NEWT:272559NEWT:3641NEWT:383379NEWT:466585NEWT:10029NEWT:1000589NEWT:85963NEWT:85962NEWT:317447NEWT:7955NEWT:7959NEWT:2261NEWT:31156NEWT:398580NEWT:4565NEWT:1264690NEWT:515619NEWT:192875NEWT:34305NEWT:59729NCBITaxon:183674NEWT:224308NEWT:84645NEWT:626528NEWT:139927NEWT:4558NEWT:209285NEWT:1283NEWT:931281NEWT:4550NEWT:1000561NEWT:197NCBITaxon:79824NEWT:4787NCBITaxon:4563NEWT:5755NEWT:44689NEWT:3218NEWT:5759NEWT:1736231NEWT:1270NEWT:2242NEWT:4784NEWT:11320NEWT:360106NEWT:286NEWT:287NEWT:10117NEWT:10239NEWT:10116NEWT:1280NEWT:1735272NEWT:83334NEWT:83332NEWT:44685NEWT:317513NEWT:1148NEWT:580240NEWT:294128NEWT:11676NEWT:55571NEWT:100226NEWT:4530NEWT:4896NEWT:75058NEWT:13616NEWT:1094343NEWT:296543NEWT:1773NEWT:1895NEWT:1182590NEWT:3712NEWT:935293NEWT:64152NEWT:4924NEWT:749200NEWT:990346NEWT:145953NEWT:257309NEWT:100816NEWT:263NEWT:230741NEWT:52283NEWT:284812NCBITaxon:1313NEWT:43330NEWT:1603293NEWT:408169NEWT:44544NEWT:4911NEWT:645463NEWT:3702NEWT:129249NEWT:243277NEWT:990119NEWT:408172NEWT:408170NEWT:493760NEWT:260710NEWT:257313NEWT:400772NEWT:3708NEWT:128161NEWT:332648NEWT:106592NEWT:536231NEWT:460519NEWT:1187947NEWT:1432138NEWT:10312NEWT:1424507NCBITaxon:1773NEWT:9598NEWT:8030NEWT:1639NEWT:188229NEWT:3818NEWT:480NEWT:4909NEWT:67767NEWT:432359NEWT:46835NEWT:1182263NEWT:2711NEWT:376686NEWT:95486NEWT:9103NEWT:29159NEWT:253NEWT:10306NCBITaxon:2759NEWT:1233435NEWT:8022NEWT:145943NCBITaxon:4932NEWT:595536NEWT:240906NEWT:593117NEWT:3635NEWT:5811NEWT:235443NEWT:272624NEWT:411483NEWT:884019NEWT:198215NEWT:411490NEWT:983964NEWT:169963NEWT:32644NEWT:499175NEWT:109779NEWT:476272NEWT:3747NEWT:195051NEWT:367830NEWT:1255228NEWT:178616NEWT:410289NEWT:373153NEWT:352472NEWT:357NEWT:360094NEWT:470NEWT:1313NEWT:411469NEWT:84023NEWT:559292NEWT:39491NCBITaxon:5811NEWT:411464NEWT:411460NEWT:2014887NEWT:2762NEWT:1174673NEWT:562NEWT:411470NEWT:33952NEWT:2094720NCBITaxon:2697049NEWT:571256NEWT:28038NEWT:1663NEWT:1423NEWT:4932NEWT:3603NEWT:2759NEWT:3847NEWT:327159NEWT:178876NEWT:327160NEWT:573NEWT:9031NEWT:7091NEWT:241368NEWT:42528NEWT:190802NEWT:9778NEWT:150475NEWT:303NEWT:9417NEWT:7111NEWT:347515NEWT:1216979NEWT:5180NEWT:256737NEWT:115104NEWT:1121114NEWT:663NEWT:1081927NEWT:1238993NEWT:67825NEWT:185579NEWT:941442NEWT:220668NEWT:13076NEWT:1249668NEWT:7108NEWT:317NEWT:7227NEWT:7469NEWT:885318NEWT:9402NEWT:415540NEWT:550NEWT:675060NEWT:4081NEWT:334542NEWT:554NEWT:98334NEWT:426428NEWT:7574NEWT:1715256NEWT:7215NEWT:575412NEWT:29204NEWT:2172103NEWT:507601NEWT:643680NCBITaxon:6157NEWT:746360NEWT:6239NEWT:470150NEWT:216257NEWT:102169NEWT:9986NEWT:4054NEWT:73239NEWT:226186NEWT:1268063NEWT:8782NEWT:1263854NEWT:435590NEWT:1902NEWT:160488NEWT:28104NEWT:1908NCBITaxon:2NEWT:985076NEWT:1215323NEWT:52641NEWT:7038NEWT:6192NEWT:28532NCBITaxon:38727NEWT:353152NEWT:2829NEWT:366581NEWT:216599NEWT:216595NEWT:51329NEWT:243230NEWT:8355NEWT:9685NEWT:7029NEWT:1080772NEWT:1283300NEWT:6183NEWT:6063NEWT:334747NEWT:61235NEWT:6289NEWT:436486NEWT:6287NEWT:300641NEWT:727NEWT:9796NEWT:725NEWT:170187NEWT:260707NCBITaxon:6191NEWT:1836NEWT:185431NEWT:29760NEWT:260704NEWT:703612NEWT:260705NEWT:80863NEWT:2697049NEWT:105231NEWT:1216981NCBITaxon:6073NEWT:884204NEWT:6279NEWT:1123869NEWT:9544NEWT:7370NEWT:83906NEWT:607699NEWT:6282NEWT:208964NEWT:1134506NEWT:575584NEWT:38783NEWT:8727NEWT:4006NEWT:8726NEWT:6426NEWT:6669NEWT:10090NEWT:4120NEWT:51515NEWT:5693NEWT:8724NEWT:51511NEWT:92867NEWT:8723NEWT:5334NEWT:37334NEWT:382352NCBITaxon:10359NEWT:242619NEWT:632957NEWT:34073NEWT:373995NEWT:5689NEWT:544404NEWT:9925NEWT:8839NEWT:4232NEWT:2758385NEWT:4113NEWT:837NEWT:11298NEWT:171101NEWT:714NEWT:36015NEWT:421932NEWT:196627NEWT:5691NEWT:48479NEWT:627025NEWT:1097677NEWT:61674NEWT:1117957NEWT:9913NEWT:4100NEWT:1076NEWT:6763NEWT:803NEWT:29722NEWT:380394NEWT:1692259NEWT:180066NEWT:135588NEWT:1843183NEWT:58002NEWT:326423NEWT:36111NEWT:4577NEWT:1416333NEWT:5664NEWT:2157NEWT:1678078NEWT:749906NEWT:418985NEWT:146479NEWT:1911079NEWT:69373NEWT:264203NEWT:1480154NEWT:1274414NEWT:27606NEWT:59202NEWT:9975NEWT:9612NEWT:38865NEWT:51953NEWT:3197NEWT:9615NEWT:10299NEWT:860688NEWT:36329NEWT:1147787NCBITaxon:3044782NEWT:72407NEWT:349741NEWT:9605NEWT:9606NEWT:157295NEWT:641501NEWT:7668NEWT:915099NEWT:74940NEWT:9721NEWT:137221NEWT:1450511NEWT:1143193NEWT:411901NEWT:9838NEWT:105884NCBITaxon:9615NEWT:9839NEWT:58334NEWT:1193501NEWT:3055NEWT:58331NEWT:6326NEWT:6689NEWT:5476NEWT:299767NEWT:1274432NEWT:1274426NEWT:70448NEWT:9825NEWT:1274423NEWT:393765NEWT:698936NEWT:39946NEWT:9823NEWT:9940NEWT:521001NEWT:39947NEWT:1274420NEWT:578458418321490000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