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Roston"],"software":["Not available"],"tissue":["Plant Cell","Shoot"],"sample_protocol":["Chloroplast Isolation and Membrane Fractionation. Chloroplasts were isolated from P. sativum as described (Bruce et al., 1994), with all buffers supplemented with protease inhibitors (1 mM PMSF, 0.5 mM ε-aminocaproic acid, and 1 mM benzamidine). Intact chloroplasts were washed and lysed by freeze-thaw and Dounce homogenization of 30 strokes using a tight pestle. Membranes were separated on a discontinuous sucrose gradient (0.46 M, 0.8 M, 1.0 M sucrose in TE buffer), followed by ultracentrifugation (180,000 × g, 3 h) as in Keegstra and Yousif (1986). IE and thylakoid fractions were recovered, diluted, and pelleted (130,000 × g, 30 min). Each was then adjusted to 0.2 M sucrose, and homogenization by freeze-thaw and Dounce was repeated before loading on a continuous sucrose gradient (0.73–1.8 M) formed by passive diffusion (Stone, 1974) and spun at 113,000 × g for 15 h. Fractions of 500 µL each were serially collected for analysis. Proteins were first precipitated with trichloroacetic acid, normalized by Bradford assay, and then loaded into the top portion of Bolt 12% Bis-Tris gels. For in-gel digestion, protein bands were excised, washed, reduced, alkylated, and digested overnight with trypsin. Peptides were extracted, dried, and resuspended in 5% acetonitrile, 0.05% trifluoroacetic acid. LC–MS/MS and Data Analysis. Peptides were analyzed by nano-LC-MS/MS using a Waters CSH C18 column online to a Thermo Orbitrap Eclipse mass spectrometer. Spectra were searched using Mascot Server (v2.7.0, www.matrixscience.com) with a fragment ion mass tolerance of 0.060 Da and a parent ion tolerance of 15.0 ppm. Variable modifications included deamidation of asparagine and glutamine, oxidation of methionine, and carbamidomethylation of cysteine."],"repository":["Pride"],"labhead":["Rebecca L Roston"],"instrument_platform":[""],"labhead_affiliation":["University of Nebraska-Lincoln"],"submission_type":["COMPLETE"],"publication":["Not available"],"submitter_affiliation":["University of Nebraska-Lincoln"],"labhead_mail":["rroston@unl.edu"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"disease":["Disease Free"],"submitter_keywords":["Heavy inner envelope","Lightthy","Inner envelope","Heavyie","Thylakoid","Light thylakoid"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD065686"],"quantification_method":[""],"modification":[""],"data_protocol":["Searches were performed against the Pisum sativum v1a annotated protein database (57,835 entries) and a common contaminants database cRAP_20150130.fasta (www.thegpm.org/crap/). Peptide and protein identifications were validated using Scaffold (v5.3.3, www.proteomesoftware.com), which was also used to calculate normalized spectral abundance factors (NSAFs). Proteins were required to have a minimum of two peptides and meet false discovery rate (FDR) thresholds of 0.1% at the peptide level and 1% at the protein level. To ensure comparability across samples, the two lowest-yielding heavyIE and lightThy samples (based on total spectral counts) were excluded from downstream analysis. The final dataset consisted of 19 samples: four inner envelope (IE), five heavyIE, four thylakoid (Thy), and six light thylakoid (lightThy) replicates. Data were processed in Perseus v2.1.3.0 (Rudolph and Cox, 2019). NSAF values were log₂-transformed, and known contaminants were removed. Protein identifications were filtered to retain only those detected in at least three biological replicates within a single sample type and at least four replicates total. Missing values were imputed from a normal distribution. Statistical comparisons were performed using a two-sided t-test modified for FDR control by permutation (Tyanova et al., 2016). Proteins were considered significantly differentially abundant if they met a log₂ fold change threshold of ±0.5 and an adjusted p-value cutoff of –log₁₀(p) ≥ 1.15 (FDR ≤ 5%)."],"omics_type":["Proteomics"],"species":["Pisum Sativum (garden Pea)"],"submitter_mail":["rroston@unl.edu"],"submitter_country":["United States"],"doi":["10.6019/PXD065686"],"additional_accession":[]},"is_claimable":false,"name":"Lipid Metabolism and Contact Site Homologs Are Enriched at the Chloroplast Envelope-Thylakoid Interface","description":"Biogenesis and maintenance of the photosynthetic thylakoid membrane requires transport of lipids from their site of synthesis in the chloroplast envelopes to their destination in the thylakoid. While vesicle trafficking is likely involved, we hypothesized a complementary mechanism involving direct membrane interactions. Using domain homology and proteomic profiling of chloroplast membrane fractions, we identified candidate lipid transport proteins enriched in a distinct, intermediate-density membrane population. This fraction was enriched in lipid metabolic enzymes and proteins homologous to known membrane organization factors. Several candidates, including TVP38 FAMILY PROTEIN (TVPFP), PLASMA MEMBRANE FUSION PROTEIN (PMFP), and LETM1-LIKE, localized to discrete subdomains within chloroplasts. Loss-of-function tvpfp or pmfp mutants exhibited altered chloroplast ultrastructure, including changes in thylakoid-envelope proximity, supporting their roles in maintaining membrane architecture. These findings identify a specialized chloroplast membrane region enriched in lipid-related functions and offer a foundation for elucidating the molecular architecture of these regions.","dates":{"publication":"2026-04-29","submission":"2025-07-02"},"accession":"PXD065686","cross_references":{"TAXONOMY":["NEWT:3555","NEWT:71647","NEWT:330879","NEWT:260201","NEWT:35554","NCBITaxon:12939","NEWT:377960","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:59753","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:1262161","NEWT:1314781","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:244366","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:1121405","NEWT:1883446","NEWT:253","NCBITaxon:2759","NEWT:1233435","NEWT:93061","NEWT:93062","NEWT:34613","NCBITaxon:4932","NEWT:1748","NEWT:109760","NEWT:29031","NEWT:108673","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:446","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:1979401","NEWT:9031","NEWT:27292","NEWT:1872122","NEWT:1308","NEWT:7091"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