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Zhang"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"software":["Not available"],"submitter_keywords":["mouse","lrrk2","proteomics","rab12 ko","striatal synaptosome","Parkinson’s disease; lrrk2; rab12 ko; mouse; proteomics;striatal synaptosome","Parkinson’s disease"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD075183"],"tissue":["Brain"],"sample_protocol":["Whole proteome profiling was performed as described previously(49, 50). Fresh lysis buffer (8 M urea, 50 mM HEPES, 0.5% sodium deoxycholate, 1× phosphatase inhibitor cocktail, pH 8.5) using for lysate striatal synaptosome (n = 5 for Rab12 KO, n = 4 for WT). 100 μL of lysis buffer was added for the striatal synaptosome pellets from each mouse. Glass beads were included to facilitate tissue homogenization using a Bullet Blender in this process. Protein concentrations were determined by BCA assay (Thermo Fisher Scientific), and 50 μg of protein from each sample was loaded onto a 10% Bis-Tris gel (BIO-RAD) and electrophoresed for 10 minutes to obtain short gel bands. Gel bands of each sample were cut and clean-up individually, followed by reduction and alkylation using DTT and iodoacetamide (IAA), respectively. Proteins were digested in-gel using trypsin at a 50:1 ratio (protein: enzyme, w:w) overnight. The supernatant from in-gel digestion was dried and resuspended in HEPES buffer (pH 8.5) for 18-plex TMTpro labeling peptides at ratio of 1:1.5 (w:w, protein: TMT reagent) for 30 minutes at room temperature. After TMT labeling, the samples were mixed equally, desalted using a Sep- Pak C18 cartridge (50mg, Waters) and dried. The pooled sample was separated on a BEH C18 column (3×150mm, 1.7μm, part NO.186004690) in 125 minutes using a gradient from 5% to 65% Buffer B at flow rate of 0.15 μL/minute (Buffer A: 10mM NH4COOH, pH 8.0; Buffer B: 10mM NH4COOH, pH 8.0, 90%ACN), and 40 fractions were collected. Each fraction was then eluted under acidic conditions at 65 °C in 95 minutes with a gradient from 8% to 65% Buffer B (Buffer A:0.2% formic acid, 3% DMSO; Buffer B: 67% ACN, 0.2% formic acid, 3% DMSO) at a flow rate of 0.25 μL/minute using a C18 column (75 μm ID* 15 cm, 1.7 μm, CoAnn) coupled to a nanoRSLC-Q-Exactive-HF system. Mass spectrometry (MS) data were acquired in positive ion mode with a resolution of 60,000 at a scan range of 460–1600 m/z for MS1 (AGC target: 1.0E6). The top 20 MS1 precursors were selected for MS2 (scan range: 200–2000 m/z; isolation window: 1.0 m/z; isolation offset: 0.2 m/z), with a maximum ion injection time of 110 ms and a dynamic exclusion of 10 seconds. Normalized collision energy (NCE) was set to 32."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["MS data search, filtering, and quantification were performed using the JUMP suite(51). For the database search, dynamic methionine oxidation (+15.99492 Da), static TMTpro modifications on peptide N-termini and lysine residues (+304.20715 Da), and cysteine alkylation (+57.02146 Da) were specified. For filtering, an initial false discovery rate (FDR) of 0.2 was applied, followed by multistep FDR filtering and fully removal of one-hit wonders. A minimum peptide length of six amino acids was required, allowing up to two missed cleavages and a Jscore threshold >10. For quantification, precursor peak intensity threshold was set at 50%, with minimum and median intensities of peptide-spectrum matches (PSMs) at 1,000 and 5,000, respectively, and minimum and median protein intensities at 2,000 and 10,000."],"omics_type":["Proteomics"],"labhead":["Junmin Peng"],"instrument_platform":[""],"labhead_affiliation":["Departments of Structural Biology and Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA"],"submission_type":["PARTIAL"],"species":["Mus Musculus (mouse)"],"submitter_mail":["xzhang56@stjude.org"],"publication":["42031745 Li X, Chen Y, Wang H, Zhang X, Guiberson NGL, Li X, Burré J, Peng J, Zhang H, Yue Z. Disruption of the LRRK2 substrate RAB12 facilitates neurotransmission and causes hyperactivity in mice. NPJ Parkinsons Dis. 2026 10.1038/s41531-026-01353-4"],"submitter_affiliation":["St. Jude Children's Research Hospital"],"submitter_country":["United States"],"pubmed_abstract":["RAB12 is a small GTPase and a validated substrate of LRRK2, a kinase genetically linked to Parkinson's disease (PD). While RAB12-LRRK2 signaling has been implicated in ciliogenesis and immune regulation, the neuronal function of RAB12 remains largely unexplored. Here, we investigated the role of RAB12 in synaptic physiology using Rab12 knockout (KO) mice. Rab12 KO mice developed normally but exhibited increased locomotor activity in adulthood. Electrophysiological recordings from striatal slices revealed enhanced presynaptic release probability and increased excitatory drive onto medium spiny neurons. Consistently, live-cell imaging of cultured cortical neurons revealed that Rab12 deletion facilitated, while Rab12 overexpression inhibited, synaptic vesicle exocytosis. Biochemical fractionation showed enrichment of RAB12 in synaptic vesicle-associated fractions containing presynaptic components. Proteomic analysis of Rab12 KO striatal synaptosomes further identified alterations in proteins involved in synaptic membrane trafficking pathways. Together, these findings establish RAB12 as a negative regulator of synaptic vesicle exocytosis and excitatory neurotransmission in vivo. Our study defines a physiological role for RAB12 in synaptic function and provides a basis for future investigation into how LRRK2-dependent RAB12 signaling may contribute to neuronal dysfunction in PD."],"pubmed_title":["Disruption of the LRRK2 substrate RAB12 facilitates neurotransmission and causes hyperactivity in mice."],"pubmed_authors":["Li Xingjian X, Chen Yuanxin Y, Wang Huaixing H, Zhang Xue X, Guiberson Noah Guy Lewis NGL, Li Xianting X, Burré Jacqueline J, Peng Junmin J, Zhang Hui H, Yue Zhenyu Z"],"additional_accession":[]},"is_claimable":false,"name":"Disruption of LRRK2 Activator RAB12 Facilitates Neurotransmission and Causes Hyperactivity in Mice","description":"RAB12 is a family member of small GTPases best known as a physiological substrate of LRRK2 kinase, the variants of which cause Parkinson’s disease (PD). RAB12 - LRRK2 interaction is crucial for the activation of LRRK2 kinase activity and pathogenic signaling in ciliogenesis and immune response. However, the function of RAB12 in neuronal activity and in vivo remains largely unexplored. Here we describe the consequence of Rab12 knock-out (KO) in mice and the role of RAB12 in regulating synaptic trafficking in neurons. Rab12 KO mice developed normally but exhibited enhanced locomotor activity in adulthood. Measuring electrophysiologic properties of Rab12 KO brain slices showed an increased release probability of presynaptic vesicles in excitatory neurons. Live-cell imaging of cortical neurons revealed that Rab12 deletion facilitated, while Rab12 overexpression inhibited, synaptic vesicle exocytosis. Further, proteomic analysis of the Rab12 KO striatum revealed the role of RAB12 in synaptic membrane transport and regulation of synaptic pathways. Collectively, our findings show that the neuronal functions of RAB12 and LRRK2 converge on the regulation of synaptic vesicle traffic, the dysfunction of which is linked to early pathological events in PD.","dates":{"publication":"2026-04-29","submission":"2026-03-04"},"accession":"PXD075183","cross_references":{"TAXONOMY":["NEWT:3555","NEWT:71647","NEWT:330879","NEWT:260201","NEWT:35554","NCBITaxon:12939","NEWT:32046","NEWT:544496","NEWT:59529","NEWT:2042546","NEWT:32049","NEWT:1392696","NEWT:259447","NEWT:295546","NEWT:45351","NEWT:445974","NEWT:43179","NEWT:180454","NEWT:5722","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:10245","NEWT:55153","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:351581","NEWT:43186","NEWT:428146","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:498019","NEWT:1351","NEWT:1438992","NEWT:1352","NEWT:2649997","NEWT:1147161","NEWT:661410","NEWT:638632","NEWT:43150","NEWT:263737","NEWT:224326","NEWT:1333499","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:5702","NEWT:95648","NEWT:1589","NEWT:135622","NEWT:1349","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:1401257","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:44491","NEWT:641809","NEWT:27933","NEWT:556484","NEWT:317447","NEWT:4688","NEWT:7955","NEWT:7719","NEWT:7959","NEWT:2261","NEWT:868565","NEWT:135674","NEWT:94323","NEWT:31156","NEWT:3112","NEWT:79329","NEWT:4442","NEWT:192875","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:284218","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:1316931","NEWT:224308","NEWT:3347","NEWT:511145","NEWT:160621","NEWT:212790","NEWT:931281","NEWT:4432","NEWT:5762","NEWT:5763","NEWT:658457","NEWT:1310161","NEWT:77133","NEWT:295358","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:2246","NEWT:2652724","NEWT:5757","NEWT:68415","NCBITaxon:4563","NEWT:5755","NEWT:44688","NEWT:44689","NEWT:44447","NEWT:752555","NEWT:498211","NEWT:5759","NEWT:347256","NEWT:1736231","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:1392","NEWT:498217","NEWT:498216","NEWT:2242","NEWT:11320","NEWT:286","NEWT:391619","NEWT:246196","NEWT:287","NEWT:246197","NEWT:145458","NEWT:633149","NEWT:1149786","NEWT:10239","NEWT:68895","NEWT:44685","NEWT:161934","NEWT:4897","NEWT:1148","NEWT:5744","NEWT:72901","NEWT:213863","NEWT:5508","NEWT:3329","NEWT:5507","NEWT:410661","NEWT:55571","NEWT:35500","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:4896","NEWT:1287689","NEWT:1390","NEWT:11557","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:294","NEWT:1773","NEWT:1182590","NEWT:3712","NEWT:82380","NEWT:3711","NEWT:935293","NEWT:375146","NEWT:2065263","NEWT:177437","NEWT:1772","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:263","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:11988","NEWT:284812","NEWT:8175","NEWT:400667","NEWT:43330","NEWT:1603293","NEWT:408169","NEWT:44664","NEWT:47946","NEWT:3702","NEWT:1245466","NEWT:243277","NEWT:1246791","NEWT:7067","NEWT:118694","NEWT:586722","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:274","NEWT:7070","NCBITaxon:287","NEWT:408172","NEWT:408170","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1436733","NEWT:460519","NEWT:1247411","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:9238","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1303443","NEWT:333760","NEWT:485","NEWT:34607","NEWT:56636","NEWT:1513458","NEWT:1233681","NCBITaxon:40559","NEWT:506599","NEWT:2853422","NEWT:81077","NEWT:84588","NEWT:1679718","NEWT:480","NEWT:65349","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:300852","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:508771","NEWT:1883446","NEWT:253","NCBITaxon:2759","NEWT:1233435","NEWT:93061","NEWT:93062","NEWT:34613","NCBITaxon:4932","NEWT:109760","NEWT:29031","NEWT:943274","NEWT:644223","NEWT:1263082","NEWT:235443","NEWT:108458","NEWT:5936","NEWT:272623","NEWT:272624","NEWT:320637","NEWT:886559","NEWT:3750","NEWT:55521","NEWT:983964","NEWT:118499","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:1130798","NEWT:55529","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:476272","NEWT:3747","NEWT:195051","NEWT:3988","NEWT:1116234","NEWT:561007","NEWT:1255228","NEWT:649908","NEWT:410289","NEWT:373153","NEWT:3983","NEWT:203696","NEWT:472","NEWT:352472","NEWT:97441","NEWT:1154756","NEWT:1071661","NEWT:1202531","NEWT:1202532","NEWT:360094","NEWT:1408223","NEWT:470","NEWT:1408224","NEWT:41364","NEWT:5911","NEWT:1313","NEWT:1218097","NEWT:1080348","NCBITaxon:50557","NEWT:1314","NEWT:39251","NEWT:39491","NEWT:212142","NCBITaxon:5811","NEWT:29491","NEWT:101841","NEWT:153481","NEWT:2014887","NEWT:1499105","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:571256","NEWT:40483","NEWT:63366","NEWT:63367","NEWT:688333","NEWT:215402","NCBITaxon:548681","NEWT:318586","NCBITaxon:5820","NEWT:1547","NEWT:272634","NEWT:198107","NEWT:9031","NEWT:27292","NEWT:1872122","NEWT:1308","NEWT:7091","NEWT:1034015","NEWT:760568","NEWT:141262","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:4087","NEWT:9778","NEWT:306","NEWT:150475","NEWT: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