{"database":"MassIVE","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://massive-ftp.ucsd.edu/v08/MSV000095252/"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"submitter":["Leonard Foster"],"full_dataset_link":["https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=d0db1f5e99a74d1aadcff18ba0250dc2"],"submitter_email":["foster@msl.ubc.ca"],"sample_protocol":[""],"repository":["MassIVE"],"file_size":["124"],"ptm_modification":["MS:1002864 - No post-translational-modifications are included in the identified peptides of this dataset"],"data_protocol":[""],"omics_type":["Proteomics"],"instrument_platform":["Bruker Daltonics instrument model"],"species":["Rattus Norvegicus (ncbitaxon:10116)"],"submitter_affiliation":["University of British Columbia"],"pubmed_abstract":["In an effort to gain insight into cellular systems impacted by neurotrophic trans-banglene (t-BG), global proteomic profiling and Western blot analyses were employed. Expression level changes in response to t-BG treatment were compared to those observed with nerve growth factor (NGF), a natural neurotrophic protein and functional analog to t-BG. Findings from these studies did not point to direct interception of NGF/TrkA signaling by t-BG. Instead, significant alterations in iron-binding and iron-regulating proteins were observed. While total iron levels showed no change across all treatments, intracellular iron measurements and mitochondrial iron measurements demonstrated lower ferrous (Fe<sup>2+</sup>) ion levels in t-BG treated cells but not in NGF treated cells. These results highlight a potential connection between iron regulation and neurotrophic activity, a relationship which has, to date, not been well studied. These results are also notable given that iron dysregulation occurs in most neurodegenerative disease settings, and that iron has been shown to facilitate protein aggregation and apoptotic mechanisms."],"pubmed_title":["Proteomic Investigation of Neurotrophic trans-Banglene Reveals Potential Link to Iron Homeostasis."],"pubmed_authors":["Gunawardana Piyumi B Wijesiri PBW, Gohil Khyati K, Moon Kyung-Mee KM, Foster Leonard J LJ, Williams Florence J FJ"],"pubmed_title_synonyms":["iron homeostasis., study"],"description_synonyms":["sodium salt, single-organism catabolic process, Rat, ammonium formate, 13C-labeled, Lysine Hydrochloride, Metabolic Concepts, NGFB, Irip, Neurologic Diseases, Lasers, SDH, magnesium formate, zinc salt, Norway Rats, NAALAdase, HOH, dmTAF[[II]]230, SDS, rat, STOP, devices, Lysine Acetate, domestic cat, Domestic Cats, Biological, 0451/09, 1, 2, C530001K22Rik, AGM4, Kinase, Metabolism Concept, Fs(3)Hor, Neurotrophic Factor, nup154, L Lysine, Health Related Quality Of Life, (R*, SET, 0244/09, iron homeostasis, Degenerative Condition, MACH 2 dental impression material, Biologic Drugs, cel, K, R, Weights, Arginine, Monohydrate DL-Arginine Acetate, Supervision, DmelCG4299, 671/2, Cognitions, haem catalase activity, Biological Medicine, LYS, Functions, scientific observation, Homo sapiens disease, RCB0009, nickel salt, AI315345, Transjugular Intrahepatic Portasystemic Shunt, dTAF[[II]]230, l(1)LB9, Biologic Products, cobalt (+2) salt, acetylation, arginine, tip, SH2-B PH domain-containing signaling mediator 1, Fs(3)Sz11, HSAN5, GIG27, sodium (4:1:1) salt, mTOR Serine-Threonine Kinase, Rattus norwegicus, magnesium salt, Felis domestica, 1316/02, BcDNA:HL08040, secretion, Technics, HRQOL, mTOR Serine-Threonine, PC-12, Logistics, ACGNAT, Tridymite, breakdown, death rate, Iron, Sodium Dodecyl Sulfate-PAGE, DI-2, LY57, I-2Dm, FU, formate, Shwachman-Diamond type metaphyseal dysplasia, Biologics, TIPSS, Fe, I-2PP1, G32, Taf250, label, Fu, ammonium (4:1) salt, l(1)16Fg, Sodium Dodecyl Sulfate-PAGEs, Administrative Technics, tlp, 4-Dimercapto-2, bs36h11.y1, pds, rac-Dithiothreitol, Biologic Pharmaceuticals, threo-1, Cleland's reagent, developmental stage, Degenerative Neurologic Disorders, Gel Electrophoresis, R*)-, SDS polyacrylamide gel electrophoresis of proteins, polyvinylxiloxane, Neurotrophins, cellular catabolism, CG7826, agua, CrAT, amorphous, DmelCG17228, Rats, Ly57, Gene Products, CG7835, Lysine, Neurite Outgrowth Factor, Technique, [SiO2], Siliziumdioxid, dTAF[[II]]250, Pulsed Lasers, thallium (+1) salt, cell, rubidium salt, Growth Associated Proteins, 4733401P19Rik, 2pp2a, CG10574, Cognitive Functions, T5E21_12., dTAF250, DmelCG6871, Continuous Wave Laser, Neurologic Degenerative Diseases, anatomical unit, Neuronal, body organ, Degenerative Neurologic Diseases, dSet, peptidos, Kb, atom, Ki, Pharmaceuticals, l(3)j12C8, measuring, DMCATHPO, TOR Serine Threonine Kinase, Quso G32, foramen, 1-(14)C-labeled, heme catalase activity, degradation, Biologic Medicines, disorders, Factor, mTOR, copper (+2) salt, Rnt, buffer, isoform CRA_b, Dithiothreitol, dTAF230, Concept, native protein, 0441/16, Rapamycin, L-Isomer Arginine, Acetate, Neurite Outgrowth, cupric formate, Felis domesticus, chemical analysis, proteomic analysis, condition, IMPACT, imprinted and ancient gene protein homolog, Death, aluminum formate, metabolism, FKBP12-Rapamycin Complex-Associated Protein, MET, DTL, Cleland's, Taf[[II]]250, AU020952, MACH-2 dental impression material, ferrum, DTT, ensemble, sodium dodecyl sulphate–polyacrylamide gel electrophoresis, Separations, (2S)-2-amino-5-guanidinopentanoic acid, Function, CG4579, SWDS, Administrative Technique, Neurologic Degenerative Conditions, ethanenitrile, CDA08, Lds, Silicon, Folate hydrolase 1, Hands, l(3)rK204, pharmacologic action, exafine, opening, Anabolism, biochemical pathways, SH2-Bb, l(3)rK137, RAFT-1, mode of action, Metabolic Process, apparatus, Kieselsaeureanhydrid, NCMe, Feature, Nerve Growth Factor 1, FKBP12-rapamycin complex-associated protein, leg, zinc formate, Cs-1, lead salt, protein, Cardiac Death, Technic, Neuronal Growth-Associated Proteins, element, Neurodegenerative Diseases, Neurite Outgrowth Factors, peptide, Techniques, ammonium tetraformate, peptido, TOR Serine-Threonine Kinase, congenital, Mammals, pathogenesis, SDS-PAGE, cellular degradation, 3.4.17.21, Arg, protein aggregate, Phenomenon, Iron-Binding Protein, Injectables, Neuronotrophic Factor, human disease, DmelCG2684, Maser, appliances, peptides, TAF-I, Natural, Administrative, Neuronal Growth Associated Proteins, hypoplasia, mAPC, Domestic, Biological Drugs, Estimated, Outgrowth Factor, Continuous Wave, IGAAD, DMPROSPER, lithium salt, Sputolysin, DmelCG10574, affinis, Run, AI316800, disease management, biotransformation, Shwachman-Bodian syndrome, Lyw-57, dihydrogen oxide, ammonium (2:1) salt, Neuro-degenerative disease, FK506-binding protein 12-rapamycin complex-associated protein 1, PLATEST, l(3)rO534, Pro, ZC2HC5, T5E21.12, Arginine Hydrochloride, Dmel_CG7826, AI836084, StF-IT-1, FK506 Binding Protein 12 Rapamycin Associated Protein 1, 4-dimercapto-2, HTATIP1, TAFII-250, Iron-Binding, 2.3.1.7, TAF250/230, Arginin, AI195249, SLP65, Rapamycin Target Protein, TAFII250, Mus norvegicus, PROS-1, PROS-2, Iron 56, Cleland, Psm, Biopharmaceuticals, pro, Neuronal Growth-Associated, lysine, Biologic Product, Progressive neurodegenerative disorder, Near-Death Experience, Mtap6, nickel (+2) salt, GAT, Dmel_CG7835, MNB, SLP-65, ammonium salt, Pteroylpoly-gamma-glutamate carboxypeptidase, Factors, polyvinyl siloxane polymer, Determination of Death, cobaltous formate, Administration and Organization, 3-Butanediol, CG4299, AW124434, Features, CG17603, Felis silvestris catus, TAF[[II]], Treatments, early, acqua, Iron Binding Protein, R*)-1, disease, inhibition of apoptosis, Q-Switched, Growth-Associated Protein, copper salt, beta Nerve Growth Factor, Measures and Weights, Neurotrophic Factors, Wasser, Laser, i2pp2a, Target of Rapamycin Proteins, Aerosil, beta-Nerve Growth Factor, CG1849, Carnitine acetylase, RAFT-1 Protein, Administrative Techniques, cesium salt, Nerve Growth Factor gamma Subunit, unsegmented paraxial mesoderm, instruments, dTAFII250, Ximpact, GCPII, PHAPII, Growth-Associated Proteins, TIPS, TOR Serine Threonine Kinases, l(3)10419, dmTAF8, Sodium Dodecyl Sulfate PAGE, Rattus sp. strain Wistar, polypeptide chain, dmTAF1, Injection, Metabolism, 14C-labeled, Pulsed, HL-VIII, N-acetylated-alpha-linked acidic dipeptidase I, anon-WO0140519.15, mTOR Serine-Threonine Kinases, silicic anhydride, DmF2, tiny, caperase activity, Rattus rattiscus, Life, TAF250, lod, study, PROS, Nervous System, l(2)10432, Rattus norvegicus, Runt, ipp2a2, Natural Product, CG7128, Acetylations, Measures, CATA, calcium formate, Mechanistic Target of Rapamycin Protein, Neurodegenerative Disorders, Administration, prod, time of survival, NGF-1beta, causality, disorder, cats, Characteristics, l(2)01501, TAF, 2-iodo-, BLNK-S, Neurotrophin, small, (SiO2)n, DYRK1, TAF[[II]]250, Nerve Growth, process quality of life cycle, protein complex, TOR Serine-Threonine, igaad, fer, l(3)84Ab, Transjugular Intrahepatic Portosystemic, medical condition, Expectancies, strontium salt, Neurotrophic, group, GCP2, Dithiotreitol, PubMed Identifier, Glutamate carboxypeptidase II, Metabolism Concepts, Axin, 2.7.11.1, Cell growth-inhibiting gene 27 protein, survival, p230, Iron Binding Proteins, I2PP2A, atomus, Dyrk1, L Arginine, CAT, ACETONITRILE, NGF receptor binding, MAP-6, optidase activity, Degenerative Conditions, nup32D, PH and SH2 domain-containing signaling mediator, silica, TAF[[II]]230, PRSS, biodegradation, Metabolic, Kinases, FKBP12-Rapamycin Associated Protein, CC1, Tripcellim, l(1)19Ea, Iron-56, CGI-97, sample population, chromic formate, Drug, Protein Gene Products, neurotrophin receptor ligand, dSET/TAF-Ibeta, 2610030F17Rik, DmelCG17603, Therapeutic, concentration, Cat01, AI425885, calcium salt, Neurodegenerative Disease, Treatment, TAF8, 1700112N14Rik, TAF1, Gunn rats, PLIP, MeCN, cadmium salt, Product, Laboratory, BOUND WATER, growth and development, End-Of-Life, CG17228, Type 2A-interacting protein, vinyl polysiloxane polymer, 1135/09, Bca, 5730420M11Rik, l(1)AA33, 1135/07, Polypeptides, NAALADase I, protein polypeptide chains, FOLH, Neurologic Disorders, cobalt(II) formate dihydrate, CH3-C#N, Dioxide, Biological Product, Mammalian target of rapamycin, 2210418N07, bacterial catalase-peroxidase activity, HTATIP, imprinted and ancient gene protein, treatment, bca, somitogenic mesoderm, Neurite, Divorced, ostium, (S)-2-amino-5-guanidinopentanoic acid, Polyacrylamide, TFIID TAF250, SDS-PAGE electrophoresis, Membrane glutamate carboxypeptidase, catabolism, CT21282, DROPROSA, metabolic process resulting in cell growth, proteins, Divorces, NTef2, AI047805, set, column, cPLA2, Degenerative Neurologic Disease, l(1)B2/13.1, Lysin, sample, Quso G-32, Medicine, Ngfb, Cristobalite, stage, RAFT 1 Protein, Biologic Drug, TIP, equilase activity, unsegmented mesenchyme, metabolism resulting in cell growth, TAF200, Insight, ANAC091, A330009E03Rik, Putative MAPK-activating protein PM10, H2O, Dm1, Life Extension, atomo, TIP60, atome, Years of Potential Life Lost, Hydrogen Oxide, Q-Switched Laser, HLA-DR-associated protein II, Degenerative Diseases, Biological Medicines, Biological Drug, FGCP, atoms, Rapamycin and FKBP12 target 1, 1167/13, cromium (+3), Pulsed Laser, Shwachman syndrome, LNKN-1, alpha, Neurologic Disease, Neurodegenerative disease, beta Trypsin, rats, Felis sylvestris catus, Pro-rich, Coordination, TAF-IBETA, CG2684, TAF-Ibeta, methyl cyanide, fused, DROCATHPO, E430016J11Rik, 0989/01, iron, lead (+2) salt, TIP41, TAF230, Cleland Reagent, FUSED, F10B6_15, knobbly, Transjugular Intrahepatic, nickel formate dihydrate, lead formate, Processes, aluminum salt, Fused, 2-Amino-5-guanidinovaleric acid, 4-dimercapto-, Metabolic Processes, protein-containing complex, 0763/13, F10B6.15, ESA1, ghenesil, Cardiac, Stable tubule-only polypeptide, manganese catalase activity, Harg, Polyacrylamide Gel Electrophoresis, disease or disorder, CG42273, TOR, Prosp, mTOR Serine Threonine Kinases, Biologicals, F15E12_6, Serine-Threonine Kinases, STOP145, beta-Trypsin, Metabolic Concept, methanoic acid, l(3)rH013, Injectable, Neurologic, Prodos, DmelCG4579, ms(2)zk, Q-Switched Lasers, 2PP2A, mechanism of action, congenital lipomatosis of pancreas, Target of Rapamycin Protein, dSET, L-Arg, Spinal Cord, Administrative Coordination, D8Wsu49e, Biologic, SDS PAGE, LB5, 0671/02, 3-butanediol, cellular breakdown, Products, 2810411E12Rik, Neurologic Disorder, TOR Kinases, Silica, BcDNA:LD21772, Proteins, Clelands Reagent, Reprosil, BG:DS00004.13, potassium formate, acetonitrile, L-Isomer, Cell, B930045J24, lLB5, polypeptide, l(3)rL433, l(3)rI160, equipment, I-2PP2A, NGF, Laboratory Rats, Dm I-2, post-mortem, Biopharmaceutical, TAF[[II]]250/230, 145-kDa STOP, segmental plate, Mopsm, Sh2bpsm1, L-Lysine, ATCMPG1, Metabolic Phenomenon, ATCMPG2, CS1, multicellular organism metabolic process, Transjugular Intrahepatic Portasystemic, lithium formate, underdeveloped, postnatal growth, NGF 1beta, Shwachman-Bodian-Diamond syndrome, CG6871, mTOR Protein, Neuronotrophic, beta, ME-IV, Neurotrophic Proteins, Cleland's Reagent, Gene Proteins, Expectancy, Pancreatic insufficiency and bone marrow dysfunction, Central Nervous System, nerve growth factor receptor ligand, mTOR Serine Threonine Kinase, Norway, beta-Nerve Growth, [OH2], neurotrophin, General activity, Enisyl, multicellular organismal catabolic process, Cognitive Function, Nup32D, deceased, IPP2A2, pharmacodynamics, Nerve growth factor, Activity, determination, DL-Arginine Acetate, postnatal development, Domestic Cat, Norway Rat, Monohydrate, Neurologic Degenerative Condition, Measure, \"Neurodegenerative disease\" EXACT [NCI2004_11_17:C4802], Neuronal Growth Associated Protein, Serine-Threonine Kinase, diseases, kinky, dJ69E11.3, Concepts, Beta-NGF, diseases and disorders, Min, 26Fe, Norway rat, FK506 Binding Protein 12-Rapamycin Associated Protein 1, beta-NGF, DmelCG42273, Nervous System Degenerative Diseases, SH2-B, FKBP12 Rapamycin Associated Protein, MUB3_18, (S)-2-Amino-5-guanidinovaleric acid, min, epsilon-diaminocaproic acid, End Of Life, pro-survival, NAALAD1, MUB3.18, l(3)j6E2, copper, Neurodegenerative Disorder, Folylpoly-gamma-glutamate carboxypeptidase, lipomatosis of pancreas, Laboratory Rat, 0320/10, SOLO DANCERS, RUN, 380, apoptosis inhibitor activity, Neurologic Degenerative Disease, Therapies, dihydridooxygen, Medicines, Catabolism, ostia, L-Arginin, catalase-peroxidase activity, Therapy, phapii, Mechanistic target of rapamycin, Process, PRO, vinyl siloxane, aqua, Q Switched Lasers, Rattus, foramina, CYS, Thermo Scientific, SiO2, AA33, Ly-57, Shunt, catalase reaction, polyvinyl siloxane, PSM, Diseases, Prostate-specific membrane antigen, presumptive somite mesoderm, and GLY protein 2, Rapamycin Protein Target, RAPT1, Mnb, L-Arginine, and GLY protein 1, Element, growth pattern, Administrative Technic, Voila, BASH, non-developmental growth, 0664/07, hydrogen-peroxide:hydrogen-peroxide oxidoreductase activity, hydrogen hydroxide, SDS polyacrylamide gel electrophoresis, negative regulation of apoptosis, TOR Kinase, Eisen, Degenerative, SR3-5, Horka, Acetamide, Fs(3)Horka, PSMA, Polypeptide, U00145, Proteomes, Platelets, other disease, Cas1, d230, SH2 domain-containing protein 1B, norvegicus, Reagent, Nerve Growth Factor alpha Subunit, Quso G 32, L Isomer, Gene, EfW1, somitomeric mesoderm, cerebral degeneration disease, formic acid, Buffer, L-(+)-arginine, FKBP12 Rapamycin Complex Associated Protein, potassium salt, Life Quality, reduced, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, Taf230, Arabidopsis NAC domain containing protein 91, zk, F15E12.6, mKIAA1299, Masers, Metabolism Phenomena, Separated, Drugs, brown rat, Portosystemic Shunt, Aerosil 380, SH2B, Taf200, hierro, 0585/13, MACH-2, Neuronal Growth-Associated Protein, breakdown of chemical, CatA, Taf1p, mGCP, Health-Related Quality Of Life, causes, eau, non-neoplastic, Cats, Catl, taf-ibeta, cromium (+3) salt, mKIAA1278, elements, 0563/18, 2-amino-5-(carbamimidamido)pentanoic acid, sodium formate, Rattus norvegicus8, Nerve Growth Factor beta Subunit, Quso, nickel formate, 3H-labeled, Life Expectancies, Felis catus, Map-6, Continuous Wave Lasers, DmelCG1849, strontium formate, total expressed protein, 2-amino-5-guanidinopentanoic acid, Medications, DL Arginine Acetate, Carnitine acetyltransferase, impact-a, Peptide, light amplification by the stimulated emission of radiation, l(3)rJ806, Metabolic Phenomena, Pros, Degenerative Neurologic Disorder, development, Insights, Characteristic, Neuronotrophic Factors, Neurotrophic Protein, natural protein, Protein, Phenomena, DL-threo-1, Hydrochloride, TFIID, Cat, cyanomethane, breakdown of molecule, Separation, death, Cas-1, Protein Target, Scales, Mammalian Target of Rapamycin, Rapamycin target protein 1, TAF[II]250, cat, Cognitive, P235, Trypure, breakdown of substance, DmelCG7128, TCV-interacting protein, Natural Products, Outgrowth Factors, Peptid, assay, AA407739, 6-diaminohexanoic acid, growth, RWDD5, PMID"],"pubmed_abstract_synonyms":["Cognitive Function, nucleocytoplasm, Nerve growth factor, ion, Activity, Nerve Growth Factor 1, PNT-P1, FON1, DmelCG17077, Western, NGFB, Neurologic Diseases, EY3-1, Pnt, Neurologic Degenerative Condition, protein, FLORAL ORGAN NUMBER 1, \"Neurodegenerative disease\" EXACT [NCI2004_11_17:C4802], Neuronal Growth-Associated Proteins, Social Controls, Neuronal Growth Associated Protein, Neurite Outgrowth Factors, protein polypeptide chains, png, Neurologic Disorders, Blottings, SUP, symptoms, Beta-NGF, Western Blotting, 26Fe, beta-NGF, D-ets-2, 3520, protein aggregate, Formal Social Controls, Neurotrophic Factor, Pnt-P1, Western blot, Nervous System Degenerative Diseases, treatment, Neuronotrophic Factor, Neurite, Degenerative Condition, Immunoblottings, Neuronal Growth Associated Proteins, Bdr, Western Blots, proteins, Super protein, Neurodegenerative Disorder, Outgrowth Factor, sp, SNAP-25, Social, TRK1, Cognitions, Degenerative Neurologic Disease, Functions, disease management, Therapies, Ngfb, Neurologic Degenerative Disease, Western Blot, TrkA, TRKA, intracellular, Neuro-degenerative disease, single organism signaling, Therapy, screening, D-Ets-2, ets94F, 0123/09, Insight, HSAN5, Iron 56, ATP synthase D chain, Ionen, Neuronal Growth-Associated, GENA70, Progressive neurodegenerative disorder, internal to cell, Tkr, Western Immunoblotting, Mitochondrial, pntP2, Factors, Pointed-P1, Iron, Degenerative Diseases, Ets94F, Control, signs, Trk-A, Neurologic Disease, Controls, Neurodegenerative disease, Treatments, Fe, ptd, PntP2, Eisen, Degenerative, PntP1, E(E2F)3D, Growth-Associated Protein, beta Nerve Growth Factor, Neurotrophic Factors, PNTP2, iron, Regulation, PNTP1, Western Blottings, beta-Nerve Growth Factor, FLORAL DEFECTIVE 10, Regulations, Western Immunoblots, 0998/12, Immunoblotting, ETS2, Nerve Growth Factor alpha Subunit, Ets2, Nerve Growth Factor gamma Subunit, Degenerative Neurologic Disorders, Gene, SUPERMAN, protein-containing complex, mitochondrial, Neurotrophins, cerebral degeneration disease, Growth-Associated Proteins, DMPOINT1A, pntegfr, 0608/07, polypeptide chain, Gene Products, C80751, Neurite Outgrowth Factor, protoplasm, hierro, Nervous System, pointed-RC, protoplast, Neuronal Growth-Associated Protein, iones, ligand, Western Immunoblottings, Blots, Growth Associated Proteins, ions, Neurodegenerative Disorders, EK3-2, Cognitive Functions, l(3)07825, Mif1, Neurologic, Neurologic Degenerative Diseases, Neuronal, NGF-1beta, Immunoblots, Degenerative Neurologic Diseases, CG17077, Spinal Cord, l(3)j1B7, p140-TrkA, MTC, Neurotrophin, Gene., Ets, Nerve Growth Factor beta Subunit, Nerve Growth, Neurologic Disorder, findings, Formal Social Control, TRK, protein complex, Proteins, fer, Factor, Synaptosomal-associated 25 kDa protein, Neurotrophic, isoform CRA_b, Cell, Degenerative Neurologic Disorder, Insights, Neuronotrophic Factors, native protein, Neurotrophic Protein, natural protein, Social Control, NGF, Neurite Outgrowth, trk, Protein, pnt-P1, pnt-P2, l(3)s118306, Degenerative Conditions, ferrum, Blot, Function, NGF 1beta, FLO10, Neurologic Degenerative Conditions, Iron-56, Cognitive, HERP, Neuronotrophic, beta, Protein Gene Products, Neurotrophic Proteins, Gene Proteins, Ets58AB, signalling process, Therapeutic, Ion, SNAP, Immunoblot, Neurodegenerative Disease, Treatment, Outgrowth Factors, Central Nervous System, regulation, beta-Nerve Growth, Western Immunoblot, General activity, POINT, CG8705"],"name_synonyms":["iron homeostasis., study"],"additional_accession":["PXD053648"]},"is_claimable":false,"name":"Proteomic Investigation of Neurotrophic trans-Banglene Reveals Potential Link to Iron Homeostasis","description":"Neurodegenerative diseases remain an area where the causative disease mechanisms, with few exceptions, are poorly understood and which are very resistant to modern therapeutic advances. Current FDA-approved medications result in modest improvements in quality of life measures and no increase in life expectancy. In this context, insight into cellular elements that dictate neuronal survival and growth versus degradation or death (the shared feature of all neurodegenerative disorders), is valuable. Molecular agents that modulate these responses can be uniquely beneficial in such inquiries because of their ease of administration, frequent robustness in biological media, and their potential for pharmacokinetic optimization or derivatization in the hands of a medicinal chemist. Further, investigation of the mechanism of action for small molecule natural products can lead to unexpected insights, such as the serendipitous discovery of the mTOR protein (mammalian target of rapamycin), a central controller of cell metabolism. We performed a global proteomic analysis of t-BG's effects in PC-12 cells and compared it to the impact of natural nerve growth factor (NGF), a neurotrophin protein which causes similar phenotypic changes in PC-12 cells. To our surprise, while we gained insight into classic pro-growth, pro-differentiation, and pro-survival pathways, we also observed changes in iron-binding proteins in the cell that were unanticipated. Since iron dyshomeostasis is an increasingly recognized element of neurodegenerative disease, particularly in early stages of Alzheimer’s and Parkinson’s disease,10 the potential connection between neurotrophic activity and altered iron homeostasis is quite notable.\n\nCell pellets were lysed in 4% SDS 100mM tris pH 6.8 and total protein concentration was estimated with BCA assay. Approximately 20 micrograms of total protein was reduced and alkylated with dithiothreitol and iodoacetamide (PMID12724530) and loaded onto 10% SDS-PAGE. Each sample's lane was separated into 5 fractionations and the proteins were digested out of the gels using MS-grade trypsin (PMID8779443). Resulting peptides were cleaned with C-18 STop And Go Extraction (STAGE) tips (PMID 12498253) using 40% (v/v) acetonitrile in 0.1% (v/v) formic acid as the elution buffer. Peptide concentration was estimated with NanoDrop One (Thermo Scientific) to load equal amounts of total peptides on Impact II Qtof (Bruker Daltonics) coupled to easy nLC 1200 (Thermo Scientific) using a in-house constructed column set up consisting a fritted 2-cm-long trap column made with 100-?m-inner diameter fused silica and 5 ?m Aqua C-18 beads (Phenomenex, cat 04A-4331) packing material, and a 30-50 cm-long analytical column with an integrated spray tip (6-8 ?m-diameter opening, pulled on a P-2000 laser puller from Sutter Instruments) made with 75-?m-inner diameter fused silica and 1.9 ?m-diameter Reprosil-Pur C-18-AQ beads (Dr. Maisch, cat r119.aq.0003) packing material. The column was heated to 50degC with another in-house constructed column heater, and was used for 60 min sample separation using the acquisition settings described in PMID 32539747. Four biological replicates of the treatment sets were processed and each sample was analyzed twice for duplicate technical replicates. The samples were randomized before injection.\nThe resulting data were searched on MaxQuant version 1.6.7.0 (PMID27809316) using Uniprot's rat proteome and common contaminants. Label-free quantitation, and match-between-run options were enabled using fixed modification of carbamidomethylation of cysteines, and variable modifications of oxidation of methionines and acetylation of protein N-termini. Specific proteolytic cleavages after arginine and lysine with up to 2 missed cleavages were set. The data were filtered for 1% false discovery at protein, peptide and PSM levels using revert decoy search mode.","dates":{"publication":"Thu Jul 04 16:36:00 BST 2024"},"accession":"MSV000095252","cross_references":{"pubmed":["40085355"]}}