Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/proteinGroups_DIA.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/proteinGroups-weiss.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/proteinGroups.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_20_WT_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_14_ZNG1mut_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_WT-5.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/WT2.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_6_ZNG1mut_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_cbwd1-KO-2.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_12_WT_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/KO1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_WT-1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_3_WT_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_18_WT_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/WT3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_21_ZNG1mut_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_24_ZNG1mut_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_7_ZNG1mut_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_WT-4.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_15_ZNG1mut_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/KO3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/KO2.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_2_WT_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_23_ZNG1mut_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_22_ZNG1mut_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_cbwd1-KO-3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_8_ZNG1mut_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_16_ZNG1mut_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_9_WT_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_WT-3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_cbwd1-KO-4.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_1_WT_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_17_WT_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_13_ZNG1mut_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_19_WT_Beng50.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_11_WT_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_WT-2.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_4_WT_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/WT1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_10_WT_TPA3.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20211208_5_ZNG1mut_no-trtmnt.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_cbwd1-KO-1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/07/PXD031516/20200721_Weiss_C57_kidL_cbwd1-KO-5.rawprimary200OKeric.skaar@vumc.orgAngela KruseMass SpectrometryAll-ion fragmentationShotgun proteomicsMouseTargeteedShotgunZinchttps://www.ebi.ac.uk/pride/archive/projects/PXD031516BrainCell CultureKidneyKidneys were isolated from 11-13-week old C57BL/6 or C57BL/6 Zng1-/- mice that were fed a low Zn diet for 5 weeks. Animals were humanely euthanized, kidneys isolated, and flash frozen in liquid nitrogen. For protein isolation, frozen organs were gently thawed on ice and transferred through a 70 µM sterile cell strainers (Thermo Fisher Scientific, 22-363-548) into 5 mL of ice-cold PBS supplemented with Halt protease inhibitor (Thermo Fisher Scientific, 78430). Filters were flushed an additional two times with PBS with protease inhibitor. Cells were pelleted (4 C, 400x g, 10 min) and pellets resuspended in 500 µL of RIPA buffer with Halt protease inhibitor (Thermo Fisher Scientific, 78430). Following rigorous vortexing, incubation on ice (10 min), and a second round of vortexing, cellular debris were pelleted by centrifugation (4 C, 10,000x g, 10 min) and supernatant transferred into fresh tubes for further analysis. Protein concentrations were determined using the 660 Protein Assay (Pierce, 22660). Equal amounts of protein (200 µg) were processed for LC-MS/MS using s-traps (Protifi) (HaileMariam et al., 2018; Zougman et al., 2014). Briefly, proteins were reduced with dithiothreitol (DTT), alkylated with iodoacetamide (IAA), acidified using phosphoric acid, and combined with s-trap loading buffer (90% MeOH, 100mM TEAB). Proteins were loaded onto s-traps, washed, and finally digested with Trypsin/Lys-C (1:100, w:w; enzyme:protein) overnight at 37 C. Peptides were eluted and dried with a vacuum concentrator. Peptides were resuspended in H2O/1% acetonitrile/0.1% formic acid for LC-MS/MS analysis. Peptides were separated using a 75 µm x 50 cm C18 reversed-phase-HPLC column (Thermo Fisher Scientific, 164570) on an Ultimate 3000 UHPLC (Thermo Fisher Scientific) with a 120 min gradient (2-32% ACN with 0.1% formic acid) and analyzed on a hybrid quadrupole-Orbitrap instrument (Q Exactive Plus, Thermo Fisher Scientific). Full MS survey scans were acquired at 70,000 resolution. The top 10 most abundant ions were selected for MS/MS analysis. Murine tissue samples: Murine brain tissue was homogenized in NP40 buffer (50mM trizma base, 150mM sodium chloride, 1mM EDTA, and 1% nonidet 40) using cycles of sonication. Protein concentration was quantified using a Bicinchoninic Acid assay (Sigma Aldrich). For each sample, 10 μg of protein was suspended in 8M urea containing 50mM ammonium bicarbonate to a final volume of 90μL. Proteins were reduced using a final concentration of 20mM dithiothreitol (DTT, Thermo Scientific) at 37°C for 45 minutes, followed by alkylation using 20mM iodoacetamide (IAA, Thermo Scientific) at room temperature for 15 minutes. Samples were diluted to reduce the concentration of urea to <1M using 50mM ammonium bicarbonate and proteins were digested using 1μg of LysN at 50°C for 2 hours. Oasis mixed-mode cation exchange (MCX) solid phase extraction cartridges (Waters Corporation, Milford, MA) were used to remove lipids and other contaminants from the digested peptides. Peptide samples were dried using a centrifugal evaporator. Mass Spectrometry Quantification of Zng1 Peptides: Dried digested samples were reconstituted in HPLC-grade water containing 0.1% formic acid by vortexing and centrifuged briefly to remove any particulates. A total of 250ng of each sample was desalted and concentrated using C18 EvoTips (EvoSep). Liquid chromatography mass spectrometry was performed using an EvoSepOne (EvoSep) coupled to an Orbitrap Fusion (ThermoFisher Scientific) equipped with a Nanospray Flex Ion source. Chromatography was performed using a 15cm long column containing ReproSil-Pur C18 1.9 µm beads by Dr Maisch C18 and a fused silica emitter tip (20 µm inner diameter) (EvoSep). Peptides were eluted from the column using an EvoSep 20 samples/day Whisper method. Mass spectrometry was performed on an Orbitrap Fusion (ThermoFisher Scientific) equipped with a Nanospray Flex Ion source. MS1 was performed using Orbitrap detection at a resolution of 60,000 with a standard AGC target. Data independent acquisition was performed using quadropole isolation with nonoverlapping windows of 16 m/z from precursor masses 400-1008 with HCD fragmentation (33% collision energy) and Orbitrap detection with a resolution of 30,000.PrideNot availableRaw data files were processed in MaxQuant (v 1.6.14.0, www.maxquant.org) and searched against the current UniProt Mus musculus protein sequences database. Search parameters included constant modification of cysteine by carbamidomethylation and the variable modifications, methionine oxidation and protein N-term acetylation. Proteins were identified using the filtering criteria of 1% protein and peptide false discovery rate. Protein intensity values were normalized using the MaxQuant LFQ function. Label free quantitation analysis was performed using Perseus (v 1.6.14.0), software developed for the analysis of -omics data. LFQ Intensity values were Log2-transformed, and then filtered to include proteins containing at least 60% valid values (reported LFQ intensities) in at least one experimental group. Finally, the missing values in the filtered dataset were replaced using the imputation function in Perseus with default parameters (Tyanova et al., 2016). Statistical analyses were carried out using the filtered and imputed protein groups file. Statistically significant changes in protein abundance are determined using Welch’s t-test p-values and z-scores. Normalized peak areas of Zng1 peptides were extracted using Skyline.ProteomicsEric P. SkaarPARTIALDepartment of Pathology, Microbiology, and Immunology;Vanderbilt Institute for Infection, Immunology, and Inflammation. Vanderbilt University Medical Center, Nashville, TN USAMus Musculus (mouse)ark253@cornell.edu35584702 Weiss A, Murdoch CC, Edmonds KA, Jordan MR, Monteith AJ, Perera YR, Rodríguez Nassif AM, Petoletti AM, Beavers WN, Munneke MJ, Drury SL, Krystofiak ES, Thalluri K, Wu H, Kruse ARS, DiMarchi RD, Caprioli RM, Spraggins JM, Chazin WJ, Giedroc DP, Skaar EP. Zn-regulated GTPase metalloprotein activator 1 modulates vertebrate zinc homeostasis. Cell. 2022 185(12):2148-2163.e27 10.1016/j.cell.2022.04.011Vanderbilt UniversityUnited StatesZinc (Zn) is an essential micronutrient and cofactor for up to 10% of proteins in living organisms. During Zn limitation, specialized enzymes called metallochaperones are predicted to allocate Zn to specific metalloproteins. This function has been putatively assigned to G3E GTPase COG0523 proteins, yet no Zn metallochaperone has been experimentally identified in any organism. Here, we functionally characterize a family of COG0523 proteins that is conserved across vertebrates. We identify Zn metalloprotease methionine aminopeptidase 1 (METAP1) as a COG0523 client, leading to the redesignation of this group of COG0523 proteins as the Zn-regulated GTPase metalloprotein activator (ZNG1) family. Using biochemical, structural, genetic, and pharmacological approaches across evolutionarily divergent models, including zebrafish and mice, we demonstrate a critical role for ZNG1 proteins in regulating cellular Zn homeostasis. Collectively, these data reveal the existence of a family of Zn metallochaperones and assign ZNG1 an important role for intracellular Zn trafficking.Zn-regulated GTPase metalloprotein activator 1 modulates vertebrate zinc homeostasis.Weiss Andy A, Murdoch Caitlin C CC, Edmonds Katherine A KA, Jordan Matthew R MR, Monteith Andrew J AJ, Perera Yasiru R YR, Rodríguez Nassif Aslin M AM, Petoletti Amber M AM, Beavers William N WN, Munneke Matthew J MJ, Drury Sydney L SL, Krystofiak Evan S ES, Thalluri Kishore K, Wu Hongwei H, Kruse Angela R S ARS, DiMarchi Richard D RD, Caprioli Richard M RM, Spraggins Jeffrey M JM, Chazin Walter J WJ, Giedroc David P DP, Skaar Eric P EPsodium salt, Water, Food Patterns, ammonium formate, Dinitrilotetraacetate, 13C-labeled, Ethylenedinitrilotetraacetic Acid, ion, Potassium EDTA, CASP-1, H(2)O, magnesium formate, X-50, zinc salt, WATER, HOH, dmTAF[[II]]230, Edetic, T22F8_160, ATGPR7, B1, 1, 2, 3, 4, Analysis, rounded, 9, nup154, (R*, Disodium Salt, C79325, Analyses, cel, N, Tissue, Basen, (24)NaCl, Processed cyclic AMP-responsive element-binding protein 3-like protein 1, D17Mit170, Edetate, HPLC, Food, isolation and purification, salt, 15-disilaoctadecane, Peptidohydrolase Inhibitor, scientific observation, infertility, D1, Acn, Solid Phase Extractions, Chromatography, s, Gallium EDTA, nickel salt, LC-MS2, dTAF[[II]]230, chronic obstructive lung disease [Ambiguous], Stars, Caspase-1 subunit p10, cobalt (+2) salt, CASP14, Tl3, tip, Tl2, not genetically inherited, Spectrum Analysis, DmelCG7788, CHRONIC OBSTRUCTIVE AIRWAY DIS, common salt, sodium (4:1:1) salt, 11-tetrathia-4, magnesium salt, Base2, Base1, IMD3, simple tissue, Chromium EDTA, FPF, Chronic obstructive lung disease, instrument, FU, NKTL, formate, Solid Phase, Sodium Salt, nitrogen, Taf250, OBSTRUCTIVE PULMONARY DISEASE (COPD), liquid chromatography-tandem mass spectroscopy, Fu, ammonium (4:1) salt, Mouse, orthophosphoric acid, Peptide Peptidohydrolase Inhibitor, Edetate Disodium Calcium, tlp, STARS, 4-Dimercapto-2, Ethylenediaminetetraacetic Acid, Old astrocyte specifically-induced substance, Gallium, rac-Dithiothreitol, Endopeptidase Inhibitors, threo-1, Ly62, DrICE, Cleland's reagent, Pierce, mini-ICE, CH3OH, CHRONIC OBSTRUCTIVE PULM DIS, precursor, Chronic obstructive pulmonary disease NOS, R*)-, TR-AP, 17alpha)-isomer, CG7826, agua, mass-to-charge ratio, Chronic obstructive pulmonary disease finding, PULMONARY DISEASE (COPD), Magnesium Disodium, isolation, Oasis, GP39, Diets, Gene Products, CG7835, Mus musculus domesticus, Technique, dyslipoproteinemic corneal dystrophy, dTAF[[II]]250, Food Pattern, GKLP, Acinus, thallium (+1) salt, cell, (22)Na, PCE-2, rubidium salt, pk18, ions, Antagonist, MBR, High-Performance Liquid Chromatographies, acinusL, Study, dTAF250, Tetracemate, Enzyme, Caspase-1 subunit p20, ATGRP7, Protease Antagonists, acinusS, ATGRP8, PTBR, Chronic, Peptide Peptidohydrolase Inhibitors, peptidos, UPLC, Kb, PTHB1, Ki, measuring, NTKL, cou, 1-(14)C-labeled, chronic obstructive airways disease NOS, Stannous, Methyl alcohol, copper (+2) salt, ACP5, COLD - Chronic obstructive lung disease, buffer, Dithiothreitol, mKIAA0670, dTAF230, Lr, MS, native protein, Temperatures, carbonyldiamide, C18, cupric formate, chemical analysis, chronic airway obstruction, NaCl, Edathamil, chronic obstructive, aluminum formate, ami, Mass Spectrum Analyses, DTL, Cleland's, Taf[[II]]250, AU020952, Calcium, DTT, Ethylene Dinitrilotetraacetate, Separations, GR-RBP7, Striated muscle activator of Rho-dependent signaling, GR-RBP8, 3-Indolylessigsaeure, CG4579, OASIS, House Mice, Monopotassium Salt, azote, ethanenitrile, Interleukin-1 beta-converting enzyme, CDA08, Magnesium Disodium EDTA, plan specification, multicellular animals, High Pressure, chronic obstructive lung disease (disorder), Peptide Hydrolase Inhibitors, reversed, T22F8.160, Meth, NCMe, Chronic Airflow Obstructions, Mus domesticus, instrument configuration, zinc formate, CG7788, crice, lead salt, Ethylenedinitrilotetraacetic, protein, peptide, Tnfsf5, Techniques, ammonium tetraformate, IAA, Uni-Etch, peptido, Edetic Acid, Chronic Obstructive Pulmonary Disease (COPD), protein aggregate, Hydrolase Inhibitors, Calcium Tetacine, wood alcohol, COLD (chronic obstructive lung disease), LCMSMS, peptides, NEC in ICD9CM_2006, E927b, 2610036I19Rik, hypoplasia, mAPC, IBP, SO, T1, Chronic airflow limitation, Tudor repeat associator with PCTAIRE-2, lithium salt, mKIAA1315, Sputolysin, ICE, GLYCINE RICH PROTEIN 7, AI316800, Tetacine, TEAB, Bladder pain syndrome, T5ap, dihydrogen oxide, MGC - 45012, ammonium (2:1) salt, So, CD40L, CHRONIC, obstructive lung disease, Calcium Disodium Edetate, iCE, drice, Nucleobase, ZC2HC5, mouse, Dmel_CG7826, Th, 4-dimercapto-2, HTATIP1, CED-4, TAFII-250, TAF250/230, Airflow Obstructions, Spectroscopy, Versenate, TAFII250, Cleland, drIce, heavy chain disease., CHRONIC OBSTRUCTIVE, Lipid, TNF-related activation protein, IES, nickel (+2) salt, Copper EDTA, Dmel_CG7835, Crystal, CHRONIC OBSTRUCTIVE LUNG DIS, MNB, absence of, ammonium salt, Painful Bladder Syndrome, Distannous, MS/MS, mice, Sonications, Encephalon, P45, cobaltous formate, AW743063, 3-Butanediol, AW124434, MICE, CG17603, TAF[[II]], Methodological Study, acqua, domesticus, R*)-1, COPD - Chronic obstructive pulmonary disease, TrATPase, IGM, copper salt, Base, Antiproteases, p45, Liquid, Wasser, 1728, Bladder Pain Syndrome, the brain, Procedures, TRAP, p50, cesium salt, dTAFII250, 2-(1-benzyl-4-methylpyrazol-3-yloxy)-2-methylpropionic acid, Spectrum Analyses, Methylalkohol, hardware, Potassium Salt, alpha-lecithin:cholesterol acyltransferase deficiency, Cold, House, dmTAF1, 14C-labeled, Mass, Studies, Protease Antagonist, Low, tiny, CD40-L, TAF250, Cold Temperatures, l(2)10432, Glass, MS1, Swiss, MS2, Indoleacetic acid, calcium formate, Xylella fastidiosa (strain Temecula1 / ATCC 700964), Chromium Salt, TEIF, wood naphtha, male sterility 1, Ultra Performance Liquid Chromatography, HIGM1, Cation, l(2)01501, TAF, Dicobalt EDTA, 2-iodo-, Disodium EDTA, C78062, High-Performance Liquid Chromatography, small, P105, COAD - Chronic obstructive airways disease, DYRK1, TAF[[II]]250, protein complex, PCTAIRE2BP, l(3)84Ab, Bp50, strontium salt, cationes, Dithiotreitol, Axin, LC/MS/MS, p230, 16-Dioxa-8, water, Metazoa, periodic FEVER, Dyrk1, CES2A1, (3alpha, ACETONITRILE, Disodium Ethylene, tandem MS, Temperature, Mass Spectrum, nup32D, with urinary tract Abnormality and cryptorchidism, TAF[[II]]230, PRSS, Chelaton 3, MALE STERILITY 1 PROTEIN, Dihydrate, CC1, Tripcellim, Disodium, chromic formate, sample population, Laboratory Mice, Protein Gene Products, Interleukin-1 beta convertase, cAMP-responsive element-binding protein 3-like protein 1, DmelCG17603, concentration, 3.4.22.36, calcium salt, cold (chronic obstructive lung disease), Chronic Obstructive Lung Disease, Chromium, 1700112N14Rik, TAF1, 3-OHECN, sodium chloride, Kation, PLIP, IL1BC, MeCN, cadmium salt, Laboratory, BOUND WATER, chronic obstructive pulmonary disease (COPD), CREB3L1, 2-(indol-3-yl)ethanoic acid, Basodexan, oxidane, House Mouse, Type 2A-interacting protein, RP11-508D10.1, Dicobalt, Polypeptides, cobalt(II) formate dihydrate, Method, Protease Inhibitor, FLASH, CH3-C#N, High Performance Liquid Chromatography, Chronic irreversible airway obstruction, GRP8, GRP7, Il1bc, HTATIP, Animalia, type 5 acid phosphatase, Divorced, TFIID TAF250, Glycine, CAFL - Chronic airflow limitation, HardwareType, Divorces, T6G21.3, RIP25, suprasegmental levels of nervous system, IL-1BC, AI047805, T-cell antigen Gp39, column, cPLA2, disease (COPD), ACN, sample, 2-Ethanediylbis(N-(carboxymethyl)glycine), High Speed Liquid, TNFSF5, TIP, suprasegmental structures, BZRP, Trypsin/K, Tnfrsf5, Dipotassium Salt, familial, Liquid Chromatography, TAF200, Procedure, ANAC091, alpha-lecithin cholesterol acyltransferase deficiency, PBS, hCD40L, PBR, Putative MAPK-activating protein PM10, H2O, (Indol-3-yl)acetate, Stickstoff, Dm1, Hydrolase Inhibitor, Magnesium Salt, TIP60, Airflow Obstruction, Hydrogen Oxide, GLYCINE-RICH PROTEIN 8, Chronic Obstructive Airways Disease, Acid, Caspase-14 subunit p10, Antagonists, Swiss Mouse, Spectrometry, cromium (+3), Caspase-14 subunit p19, LNKN-1, Methodological, N'-1, beta Trypsin, Speed, Painful bladder syndrome, heteroauxin, Dietary Patterns, F2G1.4, spirit of wood, Drice, 2018, Endopeptidase, methyl cyanide, fused, Disodium Versenate, DRICE, lead (+2) salt, TIP41, TAF230, BPBS, Cleland Reagent, FUSED, 3.4.22.-, knobbly, LCATA deficiency, nickel formate dihydrate, lead formate, Biocatalysts, DrIce, TAPK, aluminum salt, CAO - Chronic airflow obstruction, Ethylenediaminetetraacetic, Fused, 4-dimercapto-, Carbamide, protein-containing complex, abdominal muscles, thomson, animals, Chalk, Patterns, and RNA binding 1, ESA1, LC-MSMS, CG42273, Peptide Hydrolase, Carmol, Partial LCAT deficiency, gp39, CD154, CATC4, beta-Trypsin, Monosodium Salt, Extractions, methanoic acid, PCTAIRE2-binding protein, DmelCG4579, ms(2)zk, MGC:45012, Stannous EDTA, NEC, Kochsalz, D8Wsu49e, Dietary Pattern, 548, Chromatographies, Potassium, Drl, 3-butanediol, caspase 3, BcDNA:LD21772, Proteins, COAD, Kationen, Clelands Reagent, Coprin, BG:DS00004.13, potassium formate, CE-2, backward, acetonitrile, SPENCDI, encephalon, Copper, Cell, chronic, K-etchant, B930045J24, Mdu, Versene, polypeptide, Natriumchlorid, mda, TAF[[II]]250/230, 5alpha, MeOH, (COPD), eagle-Barrett syndrome, CAL - Chronic airflow limitation, lithium formate, underdeveloped, alpha-LCAT deficiency, med, IL-1 beta-converting enzyme, Peptidohydrolase Inhibitors, mDRC, ME-IV, Cleland's Reagent, Gene Proteins, Peptidase, Antiprotease, Disodium Ethylene Dinitrilotetraacetate, Bra, liquid chromatography tandem mass spectrometry, TRACP, nucleobases, Condact, [OH2], Calcium Salt, chronic obstructive pulmonary disease, liquid chromatography tandem mass spectroscopy, Nup32D, Endopeptidase Inhibitor, determination, and rna binding 2, cation, metazoans, Peptide Hydrolase Inhibitor, CASP-14, Calcium Disodium, CG11121, 10, Alkylations, 15, old, halite, BANF, kinky, dJ69E11.3, chronic obstructive airways disease NOS (disorder), Inhibitors, Min, synganglion, DmelCG42273, T-BAM, Mass Spectrum Analysis, 3-hydroxy-5-estrane-17-carbonitrile, me75, Edetates, tartrate-resistant acid ATPase, long, bases, 2610510L13Rik, min, Swiss Mice, purification, copper, Calcitetracemate, cloruro sodico, Calcium Disodium Versenate, TRAP3, fSAP152, High-Performance, Disodium Calcitetracemate, Methodological Studies, Ice, fish-eye disease, dihydridooxygen, High-Performance Liquid, glycine-rich RNA-binding protein 8, CHRONIC OBSTRUCTIVE PULMONARY DISEASE, Concise etchant, rock salt, ice, Peptidase Inhibitor, LC-MS/MS, Cd40l, aqua, Peptide Peptidohydrolase, Thermo Scientific, ur, pulmonary disease (COPD), carbamide, AA387232, HCD, Ionen, drICE, Mini-ICE, Distannous EDTA, chronic obstructive pulmonary disease and allied conditions, Chronic airway disease, NOS, Ly-62, COPD NOS, 15-tetraethoxy-, carbinol, Mnb, parent ion, Mus musculus, Chronic airway obstruction, cold, Xylella fastidiosa str. Temecula1, hydrogen hydroxide, AI326936, natrii chloridum, Chronic Airflow Obstruction, Kidneys, nanospray, H2NC(O)NH2, Peptidase Inhibitors, precursor ion, SR3-5, Acetamide, chronic obstructive airways disease, Biocatalyst, CD40LG, Polypeptide, obstructive pulmonary disease (COPD), 2-ethanediylbis(N-(carboxymethyl))-, 7N, Inhibitor, chronic obstructive lung disease, COLD, base, d230, DISEASE (COPD), Reagent, D4Ertd659e, Gene, Protease, circadian rhythm, ACINUS, EfW1, autosomal dominant, LC-MS-MS, Buffer, formic acid, Karbamid, method, potassium salt, Pulmonary Disease, reduced, DmelCG11121, Ms1, Taf230, mixt. with silica, Arabidopsis NAC domain containing protein 91, method used in an experiment, Harnstoff, Tina, zk, Animal, Mice, Separated, Mass Spectroscopy, Taf200, somda, Ethylene, iones, chlorure de sodium, Taf1p, eau, Chronic Obstructive, Pattern, Sodium Chloride, cations, FED, (indol-3-yl)acetic acid, Trap, EDTA, liquid, cromium (+3) salt, mKIAA1278, sodium formate, COPD, nickel formate, DBI, 3H-labeled, not elsewhere classified, strontium formate, Extraction, PULM DIS CHRONIC OBSTRUCTIVE, Peptide, Chronic obstructive pulmonary disease finding (finding), PKBS, Mus, Protein, DL-threo-1, TFIID, table salt, GLYCINE-RICH RNA-BINDING PROTEIN 7, Vacuums, cyanomethane, chronic obstructive airway disease, PSEC0238, Separation, uree, Dietary, Dops, Sodium chloride (NaCl), TAF[II]250, nitrogeno, Trypure, Ion, wood spirit, TCV-interacting protein, Peptid, round, High Performance Liquid, assay, SCH, Laboratory Mouseliquid chromatography tandem mass spectroscopy, LC-MS2, Mus musculus, LCMSMS, Laboratory, Swiss, mice, Mus domesticus, mouse, Swiss Mouse, LC-MS/MS, Cell Lines, House Mice, Swiss Mice, House Mouse, Cell, domesticus, Laboratory Mice, Kidneys, LC/MS/MS, House, Mus, liquid chromatography-tandem mass spectroscopy, LC-MSMS, Line, Mus musculus domesticus, liquid chromatography tandem mass spectrometry, LC-MS-MS., Mouse, house mouse, Mice, Laboratory Mouse, reniculate kidney, LinesBru, mouse protein, Cysteine Hydrochloride, E 920, 2-amino-4-(methylsulfanyl)butanoic acid, Raw, determination, experimental, AI461847, L-Zystein, L Isomer, Gene, LFQ, protein, Computer, protein-containing complex, ter, peptide, Polypeptides, L-Isomer Methionine, Methionine, peptido, Gene Products, L-Methionine, Pgi, Del(8)44H, 2-amino-3-mercaptopropanoic acid, Cystein, Software Engineering, Half-Cystine, Gpi-1, protein aggregate, Pedameth, Computer Program, Application, Svc, Open Source Softwares, C, methods, peptides, Gpi-1r, Nlk, cisteina, Gpi-1s, L Cysteine, M, Software Application, experimental section, E-920, Open, Phi, Gpi-1t, Acetylations, Open Source Software, Computer Programs and Programming, Hcys, free, Computer Software Application, set, DmelCG4216, 2-amino-4-(methylthio)butanoic acid, Tools, CYSTEINE, grupos, Bsg75C, FREE CYSTEINE, peptidos, Half Cystine, (2R)-2-amino-3-mercaptopropanoic acid, metionina, Racemethionine, NK/GPI, Applications Software, Gpi, Open Source, Computer Software, grupo, Data Set, Zinc Cysteinate, DL-Methionine, protein complex, Proteins, acetylation, 2-Amino-4-(methylthio)butyric acid, function, MF, L-Isomer, Amf, Cys, Peptide, Source Softwares, Software Tools, Tool, group, Programs, polypeptide, Program, Computer Applications, Software Tool, mOC-X, native protein, Term, Protein, chemical analysis, Computer Applications Software, Computer Applications Softwares, Methionin, NK|GPI, Hmet, Softwares, (2R)-2-amino-3-sulfanylpropanoic acid, Software, Zystein, NK, Data Base, Met, Org, ORG, Software Applications, Col4a-1, ensemble, 2-amino-3-sulfanylpropanoic acid, Source Software, Engineering, CG4216, Rest, Gpi1-r, L-2-Amino-3-mercaptopropionic acid, Gpi1-s, L-Cysteine, (R)-2-amino-3-mercaptopropanoic acid, Gpi1-t, 2-Amino-3-mercaptopropionic acid, L-Cystein, experimental procedures, Protein Gene Products, Computer Programs, Gene Proteins, Applications, Applications Softwares, label, alpha-amino-gamma-methylmercaptobutyric acid, E920, Peptid, Polypeptide, assay, variable, Liquimeth, groupe, Peptide., Bglap-rs1, Gpi1s, Gruppe, Computer Software Applicationszebra fish, Networks, GTP, 3.4.22.-, Metalloprotein, multi-cellular organism, nucleocytoplasm, Family Member, Kinship, Laboratory, Biocatalysts, Mus domesticus, Gene, mini-ICE, CASP-14, Network, cofactors, House Mouse, zinc, Guanosine Triphosphate, GTPases, zebra danio, House, Roles, Gene Products, Mus musculus domesticus, Concepts, Life Cycle, MAP1A, Cyprinus rerio, animal, Mice, D. rerios, Family Life Cycle, protoplasm, Guanosine Triphosphate Phosphohydrolases, MetAP1A, Kinship Network, enzymes, pre-mortem, AI327394, Research, Swiss, Zebrafishes, Triphosphate Phosphohydrolases, Zn, Swiss Mice, Fishes, protoplast., genetic, set, Enzyme, 30Zn, Zink, grupos, B. rerio, Role Concepts, Zebra Fishes, negative regulation of homeostatic process, Band25, species, Kinship Networks, Family Life Cycles, Family, constitutitional genetic, AW047992, GTPase, Brachydanio rerio frankei, Zn(II), Autoregulation, close to, leopard danio, Guanosine, Zebra Fish, grupo, Family Research, Guanosinetriphosphatases, body, GTP Phosphohydrolase, Proteins, familial, CASP14, mouse, whole body, zincum, enzyme activity, function, AI227039, micronutrients, predicted, gtl11, group, Concept, Family Members, near to, organism, Zn2+, Role Concept, positive regulation of homeostatic process, Mus, Protein, Danio rerio frankei, Mini-ICE, vertebrates, Metallochaperone, Role, Vertebrata, Brachydanio rerio, cinc, Vertebrate, internal to cell, Filiation, regulation of homeostatic process, MgcRacGAP, Mus musculus, Zebra, cofactor, Danio rerio, whole organism, Caspase-14 subunit p10, ensemble, Danio frankei, mice, mKIAA0094, Swiss Mouse, Rest, trace elements, House Mice, Caspase-14 subunit p19, danio, MICE, Zebra danios, Laboratory Mice, domesticus, Protein Gene Products, Phosphohydrolases, Gene Proteins, Life Cycles, Zebra danio, living, Phosphohydrolase, 1700029C17Rik, Families, approaches, Koerper, vicinity of, Biocatalyst, Micronutrient, D. rerio, inherited genetic, Mouse, mKIAA1478, Relatives, hereditary, groupe, zebrafish, Laboratory Mouse, Gruppezebra fish, Networks, GTP, 3.4.22.-, Metalloprotein, multi-cellular organism, 2-amino-4-(methylsulfanyl)butanoic acid, nucleocytoplasm, Family Member, Kinship, Laboratory, Biocatalysts, Mus domesticus, L Isomer, Gene, mini-ICE, CASP-14, Network, cofactors, House Mouse, zinc, Guanosine Triphosphate, L-Isomer Methionine, GTPases, Methionine, zebra danio, House, Roles, Gene Products, L-Methionine, Mus musculus domesticus, Concepts, Life Cycle, MAP1A, Cyprinus rerio, animal, Mice, Pedameth, D. rerios, Family Life Cycle, protoplasm, Guanosine Triphosphate Phosphohydrolases, MetAP1A, Kinship Network, enzymes, pre-mortem, AI327394, Research, Swiss, M, Zebrafishes, Triphosphate Phosphohydrolases, Zn, Swiss Mice, Fishes, protoplast., genetic, set, Enzyme, 30Zn, Zink, 2-amino-4-(methylthio)butanoic acid, grupos, B. rerio, Role Concepts, Zebra Fishes, negative regulation of homeostatic process, Band25, species, Kinship Networks, Family Life Cycles, Family, metionina, constitutitional genetic, Racemethionine, Aminopeptidase, AW047992, GTPase, Brachydanio rerio frankei, Zn(II), Autoregulation, close to, leopard danio, Guanosine, Zebra Fish, grupo, Family Research, Guanosinetriphosphatases, DL-Methionine, body, GTP Phosphohydrolase, Proteins, 2-Amino-4-(methylthio)butyric acid, familial, CASP14, mouse, whole body, zincum, enzyme activity, function, AI227039, micronutrients, L-Isomer, predicted, gtl11, group, Concept, Family Members, near to, organism, Zn2+, Role Concept, positive regulation of homeostatic process, Mus, Protein, Danio rerio frankei, Mini-ICE, vertebrates, Metallochaperone, Role, Vertebrata, Brachydanio rerio, Methionin, cinc, Vertebrate, Hmet, internal to cell, Filiation, regulation of homeostatic process, Met, MgcRacGAP, Mus musculus, Zebra, cofactor, Danio rerio, whole organism, Caspase-14 subunit p10, ensemble, Danio frankei, mice, mKIAA0094, Swiss Mouse, Rest, trace elements, House Mice, Caspase-14 subunit p19, danio, MICE, Zebra danios, Laboratory Mice, domesticus, Protein Gene Products, Phosphohydrolases, Gene Proteins, Life Cycles, Zebra danio, living, Phosphohydrolase, 1700029C17Rik, Families, approaches, Koerper, vicinity of, Biocatalyst, Micronutrient, alpha-amino-gamma-methylmercaptobutyric acid, D. rerio, inherited genetic, Mouse, mKIAA1478, Relatives, Liquimeth, hereditary, groupe, zebrafish, Laboratory Mouse, GruppePhosphohydrolases, Guanosine Triphosphate Phosphohydrolases, zinc homeostasis., MgcRacGAP, GTP, GTPases, Phosphohydrolase, Guanosine, Metalloprotein, Guanosinetriphosphatases, AI327394, GTP Phosphohydrolase, Triphosphate Phosphohydrolases, Band25, Vertebrate, mKIAA1478, AI227039, GTPase, Guanosine Triphosphate, gtl11falseMouse kidney and cell lines LC-MS/MSZinc (Zn) is an essential micronutrient and cofactor for up to 10% of proteins in living organisms. During Zn limitation, specialized enzymes called metallochaperones are predicted to allocate Zn to specific metalloproteins. This function has been putatively assigned to G3E GTPase COG0523 proteins, yet no Zn metallochaperone has been experimentally identified in any organism. Here, we functionally characterize a family of COG0523 proteins that is conserved across vertebrates. We identify Zn metalloprotease METAP1 as a COG0523 client, leading to the redesignation of this group of COG0523 proteins as Zn regulated GTPase metalloprotein activator (ZNG1) family. Using biochemical, genetic, structural, and pharmacological approaches across evolutionarily divergent models, including zebrafish and mice, we demonstrate a critical role for ZNG1 proteins in regulating cellular Zn homeostasis. Collectively, these data reveal the existence of a family of Zn metallochaperones and place ZNG1 at the center of a new paradigm for intracellular Zn trafficking.2023-07-202022-02-09PXD031516NEWT:330879NEWT:377960NEWT:1131NEWT:2042546NEWT:259447NEWT:295546NCBITaxon:2719036NEWT:1334565NEWT:625351NEWT:258594NCBITaxon:1280NEWT:112503NEWT:1129NEWT:309807NEWT:59753NEWT:89184NEWT:309800NEWT:281395NEWT:1211601NEWT:876138NEWT:44271NEWT:193516NEWT:1073882NEWT:6819NEWT:475174NEWT:111205NEWT:1117NEWT:498257NEWT:10036NEWT:1590NEWT:10034NEWT:33114NEWT:661410NEWT:638632NEWT:224326NEWT:376619NCBITaxon:79857NEWT:1096976NEWT:3765NEWT:3767NEWT:1589NEWT:135622NEWT:35786NEWT:1580NEWT:399784NEWT:96731NEWT:656064NEWT:1582NEWT:1584NEWT:36630NEWT:383379NEWT:418106NCBITaxon:12721NEWT:10029NEWT:913645NEWT:641809NEWT:1392488NEWT:38815NEWT:317447NEWT:4688NEWT:111225NEWT:7719NEWT:868565NEWT:135674NEWT:79329NEWT:30069NEWT:12637NEWT:59729NEWT:2164133NEWT:295105NEWT:108061NEWT:60711NEWT:224308NEWT:3347NEWT:160621NEWT:212790NEWT:110368NEWT:1310161NEWT:77133NEWT:145481NEWT:1310165NEWT:29058NCBITaxon:79824NEWT:1912919NEWT:410658NEWT:44688NEWT:44689NEWT:498211NEWT:347256NEWT:5518NEWT:398007NEWT:1527468NEWT:498217NEWT:498216NEWT:11320NEWT:246196NEWT:246197NEWT:145458NEWT:44685NEWT:161934NEWT:156587NEWT:1148NEWT:5508NEWT:3329NEWT:419481NEWT:5507NEWT:410661NEWT:1140NCBITaxon:2157NEWT:1143NEWT:1287689NEWT:1094343NEWT:1462472NEWT:1336795NEWT:644042NEWT:150808NEWT:1182590NEWT:3712NEWT:3711NEWT:3714NEWT:270643NEWT:2065263NEWT:177437NEWT:10418NEWT:118698NEWT:1616117NEWT:118696NEWT:34865NEWT:52283NEWT:28174NEWT:284812NEWT:8175NEWT:43330NEWT:1603293NEWT:44664NEWT:43346NEWT:3702NEWT:1245466NEWT:3704NEWT:244366NEWT:1246791NEWT:176597NEWT:118694NEWT:2850NEWT:118691NEWT:34871NEWT:33548NEWT:52299NEWT:8165NEWT:96794NEWT:3708NEWT:332648NEWT:44670NEWT:536231NEWT:376219NEWT:219813NEWT:1510NEWT:460519NEWT:1515NEWT:572307NEWT:1432138NEWT:1424507NEWT:1194599NEWT:272844NEWT:1348799NEWT:33995NEWT:55784NEWT:1303443NEWT:9483NEWT:485NEWT:56636NEWT:2709072NEWT:2499525NEWT:2853422NEWT:487NEWT:1538553NEWT:1679718NEWT:480NEWT:67767NEWT:46835NEWT:109757NEWT:582580NEWT:1185650NEWT:216778NEWT:294607NEWT:1502NEWT:128017NEWT:376686NEWT:237610NEWT:95486NEWT:1883446NEWT:1233435NEWT:118223NEWT:11757NEWT:987059NEWT:109760NEWT:29031NEWT:317047NEWT:491NEWT:460523NEWT:45954NEWT:235443NEWT:1757312NEWT:85694NEWT:109786NEWT:108458NEWT:5936NEWT:320637NEWT:1078283NEWT:3750NEWT:983964NEWT:11706NEWT:32644NEWT:527796NEWT:499175NEWT:109779NEWT:3745NEWT:1715989NCBITaxon:4751NEWT:3747NEWT:1116234NEWT:1255228NEWT:410289NEWT:373153NEWT:1640278NEWT:472NEWT:940825NEWT:529507NEWT:1071661NEWT:28128NEWT:470NEWT:5911NEWT:748693NCBITaxon:50557NEWT:39251NEWT:29491NEWT:101841NEWT:446NEWT:153481NEWT:2014887NEWT:33952NEWT:445NEWT:153009NEWT:261756NEWT:166313NEWT:63366NEWT:63367NEWT:215402NEWT:1547NEWT:906914NEWT:28141NEWT:9031NEWT:27292NEWT:1050722NEWT:3728NEWT:108931NEWT:1293497NEWT:1055524NEWT:150475NEWT:267872NEWT:381666NEWT:172269NEWT:9534NEWT:5180NEWT:256737NEWT:9541NEWT:8694NEWT:33936NEWT:8692NEWT:2903NEWT:185579NEWT:13076NEWT:207340NEWT:1006581NEWT:8673NEWT:33940NEWT:550NEWT:554NEWT:451516NEWT:552NEWT:1325291NEWT:42410NEWT:36185NEWT:1054211NEWT:1225786NEWT:771870NEWT:51338NEWT:575412NEWT:28112NEWT:6493NEWT:10832NEWT:6494NEWT:6491NEWT:507601NEWT:520NEWT:186441NEWT:643680NEWT:214092NCBITaxon:6157NEWT:13095NEWT:162425NEWT:104105NEWT:216257NEWT:9986NEWT:8654NEWT:8658NEWT:1268063NEWT:8657NEWT:9983NEWT:8655NEWT:5147NEWT:595495NEWT:28108NEWT:28104NEWT:407821NEWT:568703NCBITaxon:2NEWT:568708NEWT:1613NEWT:986NEWT:52641NEWT:28532NEWT:353152NEWT:298176NEWT:353153NEWT:227377NEWT:1774284NEWT:40674NEWT:1194669NEWT:51329NEWT:443906NEWT:519NEWT:2510939NEWT:7394NEWT:6063NEWT:1328388NEWT:105487NEWT:1548728NEWT:667127NEWT:9557NEWT:377586NEWT:300641NEWT:39655NEWT:9554NEWT:38323NEWT:475932NEWT:256318NEWT:206411NCBITaxon:6191NEWT:229533NEWT:214053NEWT:28550NEWT:80863NEWT:90675NEWT:52639NEWT:52638NEWT:57075NEWT:1666905NEWT:8697NEWT:861109NEWT:8695NEWT:884204NEWT:510516NEWT:1045775NEWT:1123869NEWT:9544NEWT:9545NEWT:979NEWT:7370NEWT:83906NEWT:99488NEWT:1134506NEWT:255470NEWT:38783NEWT:6426NEWT:33090NEWT:9935NEWT:287889NEWT:305959NEWT:1028307NEWT:13489NEWT:29829NEWT:92867NEWT:92866NCBITaxon:3055NEWT:51750NEWT:202950NEWT:441772NEWT:1293091NEWT:48106NEWT:202956NEWT:295027NCBITaxon:11320NEWT:632957NEWT:9925NCBITaxon:9606NEWT:90690NEWT:1436183NEWT:300273NEWT:4232NEWT:416348NEWT:11298NEWT:4236NEWT:7742NEWT:196627NEWT:1076878NEWT:200308NEWT:200302NEWT:870435NEWT:58370NEWT:9913NEWT:9915NEWT:105841NEWT:2666255NEWT:999810NCBITaxon:5693NEWT:28995NEWT:2144189NEWT:1392998NEWT:380394NEWT:1150621NEWT:226900NEWT:1266738NEWT:231490NEWT:244704NEWT:7725NEWT:9909NEWT:9902NEWT:430615NEWT:563041NEWT:72664NEWT:36599NEWT:41949NEWT:249161NEWT:326423NEWT:452467NEWT:198822NEWT:36111NEWT:326424NEWT:1678078NEWT:749906NEWT:418985NEWT:749907NEWT:150847NEWT:29835NEWT:431947NEWT:69014NEWT:142809NEWT:302409NEWT:130821NEWT:27606NEWT:1519788NEWT:59202NEWT:9975NEWT:8643NEWT:1801631NEWT:1159899NEWT:502780NEWT:860688NEWT:13443NEWT:35231NEWT:1774208NEWT:8644NEWT:1125630NEWT:5141NEWT:46360NCBITaxon:3044782NEWT:20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