Lipopolysaccharide Upregulates Palmitoylated Enzymes of the Phosphatidylinositol Cycle: An Insight from Proteomic Studies

• DOI: 10.1074/mcp.ra117.000050
• Publication Date: 2017-12-08T01:50:17Z
• Authors: Justyna Sobocińska; Paula Roszczenko-...; Monika Zaręba-Kozioł; Aneta Hromada-Jud...; Orest V. Matveichuk; Gabriela Traczyk; Katarzyna Łukasiuk; Katarzyna Kwiatko...
• Keywords: Proinflammatory cytokine; Palmitoylation

Lipopolysaccharide (LPS) is a component of the outer membrane Gram-negative bacteria that induces strong proinflammatory reactions mammals. These processes are triggered upon sequential binding LPS to CD14, GPI-linked plasma raft protein, and TLR4/MD2 receptor complex. We have found earlier binding, CD14 triggers generation phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], lipid controlling subsequent cytokine production. Here we show stimulation RAW264 macrophage-like cells with global changes level fatty-acylated, most likely palmitoylated, proteins. Among acylated proteins were up-regulated in those conditions several enzymes cycle. Global profiling was performed by metabolic labeling 17ODYA, an analogue palmitic acid functionalized alkyne group, followed detection enrichment labeled using biotin-azide/streptavidin their identification mass spectrometry. This proteomic approach revealed 154 fatty-acylated up-regulated, 186 downregulated, 306 not affected stimulated 100 ng/ml for 60 min. The involved diverse biological functions, as Ingenuity Pathway Analysis. Detailed studies 17ODYA-labeled immunoprecipitated S-palmitoylation, hence activation, type II 4-kinase (PI4KII) β, which phosphorylates 4-monophosphate, PI(4,5)P2 precursor. Silencing PI4KIIβ PI4KIIα inhibited LPS-induced expression production cytokines, especially TRIF-dependent signaling pathway TLR4. Reciprocally, this significantly enhanced after overexpression or PI4KIIα; dependent on palmitoylation kinases. However, S-palmitoylation PI4KIIα, its activity, constitutive cells. Taken together data indicate activation PI4KIIβ, generates PI(4)P pathways cytokines. bacteria. Upon infection, responses facilitate eradication recognition Toll-like 4 (TLR4) (1.Poltorak A. He X. Smirnova I. Liu M.Y. Van Huffel C. Du Birdwell D. Alejos E. Silva M. Galanos Freudenberg Ricciardi-Castagnoli P. Layton B. Beutler Defective C3H/HeJ C57BL/10ScCr mice: Mutations Tlr4 gene.Science. 1998; 282: 2085-2088Crossref PubMed Scopus (6421) Google Scholar), expressed myeloid lineage, such monocytes, macrophages, dendritic cells, certain nonimmune like endothelial epithelial TLR4 belongs so-called pattern recognizing receptors specialized evolutionarily conserved constituents cell wall/membranes microbes, RNA, unmethylated CpG DNA motifs. trigger innate immune constituting first line defense against pathogens initiate appropriate adaptive (2.Brubaker S.W. Bonham K.S. Zanoni Kagan J.C. Innate recognition: A perspective.Annu. Rev. Immunol. 2015; 33: 257-290Crossref (878) Scholar). reaction, although initially beneficial, when exaggerated can lead potentially fatal systemic inflammation called sepsis, severe septic shock (3.Angus D.C. van der Poll T. Severe sepsis shock.N. Engl. J. Med. 2013; 369: 840-851Crossref (1837) In addition, prolonged low-grade caused derived from intestinal has been recognized factor development chronic diseases (4.Kaliannan K. Wang Li X.Y. Kim K.J. Kang J.X. host-microbiome interaction mediates opposing effects omega-6 omega-3 fatty acids endotoxemia.Sci. Rep. 5: 11276Crossref (206) additionally drives interest TLR4-induced signaling. transmembrane protein associates extracellular MD2 crucial binding. up five six acyl chains buried within hydrophobic pocket one while remaining chain interacts neighboring complex, inducing dimerization complexes (5.Park B.S. Song D.H. H.M. Choi Lee H. J.O. structural basis lipopolysaccharide TLR4-MD-2 complex.Nature. 2009; 458: 1191-1195Crossref (1591) facilitates recruitment two sets adaptor Toll/interleukin-1 (TIR) homology domain First, pair consisting TIR domain-containing (TIRAP) MyD88 binds receptor, guided simultaneous TIRAP both lipid, [PI(4,5)P2] (6.Kagan Medzhitov R. Phosphoinositide-mediated controls signaling.Cell. 2006; 125: 943-955Abstract Full Text PDF (666) MyD88, turn, recruits interleukin-1 receptor-associated kinase (IRAK)4 IRAK1/2, forming multimolecular complex myddosome (7.Motshwene P.G. Moncrieffe M.C. Grossmann J.G. Kao Ayaluru Sandercock A.M. Carol V. Robinson C.V. Latz Gay N.J. An oligomeric platform formed signal transducers IRAK-4.J. Biol. Chem. 284: 25404-25411Abstract (259) assembly initiates cascade, eventually leading NFκB activator protein-1 transcription factors, latter acting downstream MAP Expression genes encoding tumor necrosis α (TNFα), follows (8.Kawai Akira S. crosstalk other infection immunity.Immunity. 2011; 34: 637-650Abstract (2589) Subsequently, internalized, at endosomal membrane, set replaced another interferon-β (TRIF) TRIF-related molecule (TRAM) (9.Husebye Halaas Ø. Stenmark Tunheim G. Sandanger Bogen Brech Espevik Endocytic regulate link immunity.EMBO 25: 683-692Crossref (355) Scholar, 10.Kagan Su Horng Chow TRAM couples endocytosis induction interferon-β.Nat. 2008; 9: 361-368Crossref (947) 11.Enokizono Y. Kumeta Funami Horiuchi Sarmiento Yamashita Standley D.M. Matsumoto Seya Inagaki F. Structures interface mapping molecules interferon signaling.Proc. Natl. Acad. Sci. U.S.A. 110: 19908-19913Crossref (50) thereby leads IRF3/7 factors I interferons some cytokines exemplified C-C motif chemokine ligand 5/regulated normal T secreted (CCL5/RANTES) assisted CD14. contains glycosylphosphatidylinositol (GPI) moiety attached C terminus, anchors leaflet nanodomains rich cholesterol lipids saturated (mainly sphingolipids), rafts (12.Simmons D.L. Tan Tenen D.G. Nicholson-Weller Seed Monocyte antigen phospholipid anchored protein.Blood. 1989; 73: 284-289Crossref 13.Płóciennikowska Hromada-Judycka Borzęcka Kwiatkowska Co-operation pro-inflammatory Mol. Life 72: 557-581Crossref (409) Once large located N-terminal part facilities transfer onto (14.Kim J.-I. C.J. Jin M.S. C.H. Paik S.G. Crystal structure implications signaling.J. 2005; 280: 11347-11351Abstract (211) 15.Gioannini T.L. Teghanemt Zhang Levis E.N. Weiss J.P. Monomeric endotoxin:protein essential TLR4-dependent activation.J. Endotoxin Res. 11: 117-123Crossref growing body evidence indicates role goes beyond LPS. internalization LPS-activated TLR4/MD2, exceptions, CD14-controlled uptake required initiation (16.Jiang Z. Georgel Shamel L. Sovath Mudd Huber Kalis Keck MyD88-independent signaling.Nat. 6: 565-570Crossref (522) 17.Husebye Aune M.H. Stenvik Samstad Skjeldal O. Nilsen Lein Mollnes T.E. Bakke Rab11a GTPase 4-induced regulatory factor-3 phagosomes.Immunity. 2010; 583-596Abstract (161) 18.Zanoni Ostuni Marek L.R. Barresi Barbalat Barton G.M. Granucci 4.Cell. 147: 868-880Abstract (618) 19.Tan Cullen T.W. Goodman A.L. Mechanisms reveal common immune-evasion strategy used pathogenic commensal bacteria.Immunity. 43: 909-922Abstract (102) Our recent shown direct between LPS-stimulated LPS, biphasic accumulation whereas fine tune process (20.Płóciennikowska Zdioruk M.I. Traczyk Świątkowska clustering PI(4,5)P2.J. Cell 128: 4096-4111Crossref (20) 21.Płóciennikowska Dembińska Roszczenko Ciesielska Contribution cells.J. Leukoc. 2016; 100: 1363-1373Crossref (18) generated phosphorylation 4-monophosphate [PI(4)P] 4-phosphate 5-kinase (PIP5KI), isoforms Iα Iγ 22.Nguyen T.T. Y.M. T.D. Le O.T. J.J. H.C. Hasegawa Kanaho Jou S.Y. Phosphatidylinositol 4-mediated microglial through regulation location.J. 288: 5645-5659Abstract (38) serves site MyD88-dependent undergoes hydrolysis diacylglycerol inositol 1,4,5-trisphosphate (IP3) or, alternatively, 3,4,5-trisphosphate [PI(3,4,5)P3]. Both (18.Zanoni 23.Chiang C.-Y. Veckman Limmer David Phospholipase Cγ-2 intracellular calcium lipopolysaccharide-induced 3 (IRF3) 2012; 287: 3704-3709Abstract (93) 24.Aksoy Taboubi Torres Delbauve Hachani Whitehead M.A. Pearce W.P. Berenjeno I.M. Nock Filloux Beyaert Flamand Vanhaesebroeck p110δ isoform PI(3)K subcellular compartmentalization protects endotoxic shock.Nat. 13: 1045-1054Crossref (141) Additionally, also control TLR4-independent entered cytosol (25.Ding W. She Sun Q. Shi Shao Pore-forming activity autoinhibition gasdermin family.Nature. 535: 111-116Crossref (1249) 26.Liu Ruan Pan Magupalli V.G. Wu Lieberman Inflammasome-activated D causes pyroptosis pores.Nature. 153-158Crossref (1488) point phosphatidylinositols affecting all During CD14- TLR4/MD2-mediated newly accumulates Triton X-100-resistant fraction enriched components, including itself corroborate thesis location machinery engaged (13.Płóciennikowska One key association S-acylation, frequently referred defined attachment (C16:0) residue cysteine thioester bond (27.Levental Lingwood Grzybek Coskun U. Simons Palmitoylation regulates affinity majority integral proteins.Proc. 107: 22050-22054Crossref (380) general, more broadly, affects diversity stability, cellular trafficking, localization, protein–protein interactions (28.Chamberlain L.H. Shipston M.J. physiology S-acylation.Physiol. 95: 341-376Crossref (202) 29.Sobocińska Roszczenko-Jasińska Protein virus pathogenicity.Frontiers Immunology. 2018; (10.3389/fimmu.2017.02003)PubMed functions be disturbed when, instead acid, dietary unsaturated via linkage proteins; however, consequences S-acylation remain mostly unknown (30.Liang Nazarian Erdjument-Bromage Bornmann Tempst Resh M.D. Heterogeneous acylation Src family kinases polyunsaturated localization transduction.J. 2001; 276: 30987-30994Abstract (183) 31.Thinon Percher Hang Bioorthogonal chemical reporters monitoring proteins.ChemBioChem. 17: 1800-1803Crossref (17) Beside modifications consist residues myristoylation, lysine side acylation, rare O- N-palmitoylation (32.Resh Fatty proteins: long short it.Prog. Lipid 63: 120-131Crossref (167) these acylations, best characterized reversible modification, 24 palmitoyl acyltransferases (zinc finger DHHC (zDHHC) containing enzyme family) depalmitoylating identified mammals (33.Ohno Kihara Sano Igarashi Intracellular tissue-specific distribution human yeast cysteine-rich proteins.Biochim. Biophys. Acta. 1761: 474-483Crossref (319) 34.Blaskovic Adibekian Blanc Goot G.F. Mechanistic thereof.Chem. Phys. Lipids. 2014; 180: 44-52Crossref (35) 35.Yokoi N. Fukata Sekiya Murakami Kobayashi Identification PSD-95 enzymes.J. Neurosci. 36: 6431-6644Crossref (135) It engagement than (36.Zhang M.M. Tsou L.K. Charron Raghavan A.S. Tandem fluorescence imaging dynamic turnover.Proc. 8627-8632Crossref (87) 37.Jia Chisari Maktabi Sobieski Zhou Konopko Martin B.R. Mennerick S.J. Blumer mechanism regulating G protein-coupled requires cycles depalmitoylation.J. 289: 6249-6257Abstract (31) 38.Akimzhanov Boehning Rapid transient tyrosine Lck Fas 112: 11876-11880Crossref (56) S-palmitoylated Lyn raft-enriched determines negative (39.Borzęcka-Solarz Ziemlińska Association influence signaling.Mol. Cell. 2017; 28: 1147-1159Crossref Taking into account redistribution MyD88- TLR4, assumed could contribute substantially cascades events. order identify study metabolically analogue, 17-octadecynoic (17ODYA) 1The abbreviations are: acid; AP-1, protein-1; BPA, bromohexadecanoic BSA, bovine serum albumin; CCL5/RANTES, secreted; DGKε, kinase-ε; eIF5A2, eukaryotic translation 5A2; FBS, fetal serum; GPI, glycosylphosphatidylinositol; IP3, 1,4,5-trisphosphate; IPA, Analysis; ISRE, interferon-stimulated response element; lipopolysaccharide; PBS, phosphate buffered saline; PI, phosphatidylinositol; PI(4)P, 4-monophosphate; PI(4,5)P2, 4,5-bisphosphate; PI(3,4,5)P3, 3,4,5-trisphosphate; PI4KII, 4-kinase; PIP5KI, 5-kinase; TBTA, Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine; TCEP, Tris(2-carboxyethyl)phosphine hydrochloride; TIR, receptor; 4; VSVG, vesicular stomatitis G-protein; zDHHC, zinc DHHC; emPAI, exponentially modified abundance index; FDR, false discovery rate; IRAK, kinase; IRF, factor; TIRAP, protein; TNFα, α; TRAM, molecule; TRIF, adapter-inducing interferon-β; qPCR, quantitative polymerase reaction. then tagged biotin-azide click chemistry reaction analyzed them Proteomic analysis recently DC2.4 (31.Thinon 40.Merrick B.A. Dhungana Williams Aloor Peddada Tomer K.B. Fessler M.B. S-acylated macrophage identifies mitochondrial targeting scramblase 3.Mol. Proteomics. 10Abstract (49) 41.Chesarino N.M. Hach Chen J.L. Zaro B.W. Rajaram M.V. Turner Schlesinger L.S. Pratt M.R. Yount J.S. Chemoproteomics reveals acylation.BMC 12: 91Crossref (45) should emphasized, none examined palmitoylation, thus contribution palmitoylated remains largely unknown. downregulated ones min, time window where events related occur. Analysis (IPA) functional networks involving By adding immunoprecipitation step above procedure, confirmed pointing PI(4)P. precursor cascade RAW264.7 J774A.1 HEK293 cultured DMEM 10% (FBS), 2 mm l-glutamine, U/ml penicillin, μg/ml streptomycin. Metabolic 17ODYA (Sigma-Aldrich, Poznan, Poland) 2% charcoal-stripped FBS (ThermoFisher Scientific, Waltham, MA, USA), antibiotics 30 Hepes, pH 7.4. For stimulation, exposed (ultrapure smooth Escherichia. coli O111:B4, List Biological Laboratories, Campbell, CA, USA). experiments, prior incubated 125–250 μm (BPA) 1 h (37 °C) DMEM/2% drug present during BPA precomplexed fatty-acid-free albumin (BSA; Sigma-Aldrich) 4:1 molar ratio, essentially described (42.Kwiatkowska Frey Sobota Phosphorylation FcγRIIA receptor-induced actin rearrangement capping: rafts.J. 2003; 116: 537-550Crossref (90) When indicated, overnight 150–500 prepared according (43.Jin Lu Perry Russo S.B. Cowart L.A. Hannun Y.A. Huang Acid sphingomyelinase plays acid-amplified inflammatory low concentrations macrophages.Am. Physiol. Endocrinol. Metab. 305: E853-E867Crossref (61) min °C), presence. pCMV5-Myc plasmid rat deletion mutant lacking