Journal of Biological Chemistry (J BIOL CHEM)

Publications in this journal

• Article: A heme-based redox sensor in the methanogenic archaeon Methanosarcina acetivorans

  Bastian Molitor, Marc Stassen, Anuja Modi, Samir F. El-Mashtoly, Christoph Laurich, Wolfgang Lubitz, John H. Dawson, Michael Rother, Nicole Frankenberg-Dinkel
  Journal of Biological Chemistry 05/2013;
• Article: The choline-binding protein PspC of Streptococcus pneumoniae interacts with the C-terminal heparin-binding domain of vitronectin.

  Voss S, Hallstroem T, Saleh M, Burchhardt G, Pribyl T, Singh B, Riesbeck K, Zipfel PF, Hammerschmidt S
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  ABSTRACT: Adherence of Streptococcus pneumoniae is directly mediated by interactions of adhesins with eukaryotic cellular receptors or indirectly by exploiting matrix and serum proteins as molecular bridges. Pneumococci engage vitronectin, the human adhesive glycoprotein and complement inhibitor, to facilitate attachment to epithelial cells of the mucosal cavity, thereby modulating host cell signaling. In this study, we identified PspC as vitronectin-binding protein interacting with the C-terminal heparin-binding domain of vitronectin. PspC is a multifunctional surface-exposed choline-binding protein displaying various adhesive properties. Vitronectin-binding required the R domains in the mature PspC protein, which are also essential for the interaction with the ectodomain of the polymeric immunoglobulin receptor and secretory IgA. Consequently, secretory IgA competitively inhibited binding of vitronectin to purified PspC and to PspC-expressing pneumococci. In contrast, Factor H, that binds to the N-terminal part of mature PspC molecules, did not interfere with the PspC-vitronectin interaction. Using a series of vitronectin peptides, the C-terminal heparin-binding domain was shown to be essential for the interaction of soluble vitronectin with PspC. Binding experiments with immobilized vitronectin suggested a region N-terminally to the identified HBD as additional binding region for PspC, suggesting that soluble, immobilized as well as cellularly bound vitronectin possess different conformations. Finally, vitronectin bound to PspC was functionally active and inhibited the deposition of the terminal complement complex. In conclusion, this study identifies and characterizes (on the molecular level) the interaction between the pneumococcal adhesin PspC and the human glycoprotein vitronectin.
  Journal of Biological Chemistry 04/2013;
• Article: Physical and Functional Interactions of a Monothiol Glutaredoxin and an Iron Sulfur Cluster Carrier Protein with the Sulfur-Donating Radical S-adenosyl-L-methionine Enzyme MiaB

  Sylvain Boutigny, Avneesh Saini, Edward E.K. Baidoo, Natasha Yeung, Jay D. Keasling, Gareth Butland
  Journal of Biological Chemistry 03/2013;
• Article: Drug uptake, lipid rafts and vesicle trafficking modulate resistance to an anticancer lysophosphatidylcholine analogue in yeast.

  Alvaro Cuesta-Marbán, Javier Botet, Ola Czyz, Luis M Cacharro, Consuelo Gajate, Valentín Hornillos, Javier Delgado, Hui Zhang, Francisco Amat-Guerri, A Ulises Acuña, Christopher R McMaster, Jose Luis Revuelta, Vanina Zaremberg, Faustino Mollinedo
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  ABSTRACT: The ether-phospholipid edelfosine, a prototype antitumor lipid (ATL), kills yeast cells and selectively kills several cancer cell types. To gain insight into its mechanism of action, we performed chemogenomic screens in the Saccharomyces cerevisiae gene-deletion strain collection, identifying edelfosine-resistant mutants. LEM3, AGP2 and DOC1 genes were required for drug uptake. Edelfosine displaced the essential proton pump Pma1p from rafts, inducing its internalization into the vacuole. Additional ATLs, including miltefosine and perifosine also displaced Pma1p from rafts to the vacuole, suggesting that this process is a major hallmark of ATL cytotoxicity in yeast. Radioactive and synthetic fluorescent edelfosine analogues accumulated in yeast plasma membrane rafts and subsequently the endoplasmic reticulum. Although both edelfosine and Pma1p were initially located at membrane rafts, internalization of the drug towards endoplasmic reticulum and Pma1p to the vacuole followed different routes. Drug internalization was not dependent on endocytosis and was not critical for yeast cytotoxicity. However, mutants affecting endocytosis, vesicle sorting or trafficking to the vacuole including the retromer and ESCRT complexes, prevented Pma1p internalization and were edelfosine-resistant. Our data suggest that edelfosine-induced cytotoxicity involves raft reorganization, and retromer- and ESCRT-mediated vesicular transport and degradation of essential raft proteins leading to cell death. Cytotoxicity of ATLs is mainly dependent on the changes they induce in plasma membrane raft-located proteins that lead to their internalization and subsequent degradation. Edelfosine toxicity can be circumvented by inactivating genes that then result in the recycling of internalized cell-surface proteins back to the plasma membrane.
  Journal of Biological Chemistry 03/2013; 288(12):8405-8418.
• Article: Chaperone peptides of α-crystallin inhibit epithelial cell apoptosis, protein insolubilization and opacification in experimental cataracts

  Rooban B. Nahomi, Benlian Wang, Cibin T. Raghavan, Oliver Voss, Andrea Doseff, Puttur Santhoshkumar, Ram H. Nagaraj
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  ABSTRACT: α-Crystallin is a member of the small heat-shock protein (sHSP) family and consists of two subunits, αA- and αB-. Both αA- and αB-crystallin act as chaperones and anti-apoptotic proteins. Previous studies have identified the peptide 70KFVIFLDVKHFSPEDLTVK88 in αA-crystallin and the peptide 73DRFSVNLDVKHFSPEELKVK92 in αB-crystallin as mini-chaperones. In the human lens, lysine70 (K70) of αA- and K92 of αB- (in the mini-chaperone sequences) are acetylated. In this study, we investigated the cellular effects of the unmodified and acetyl mini-chaperones. The αA- and αB-crystallin peptides inhibited stress-induced aggregation of four client proteins, and the acetyl peptides were more effective than the native peptides against three of the client proteins. Both the acetyl and native crystallin peptides inhibited stress-induced apoptosis in two mammalian cell types, and this property was directly related to the inhibition of cytochrome-C release from mitochondria and the activity of caspase-3 and -9. In organ-cultured rat lenses, the peptides inhibited calcimycin-induced epithelial cell apoptosis. Intraperitoneal injection of the peptides inhibited cataract development in selenite-treated rats, which was accompanied by inhibition of oxidative stress, protein insolubilization and caspase activity in the lens. These inhibitory effects were more pronounced for acetyl peptides than native peptides. A scrambled αA-crystallin peptide produced no such effects. The results suggest that the α-crystallin chaperone peptides could be used as therapeutic agents to treat cataracts and diseases in which protein aggregation and apoptosis are contributing factors.
  Journal of Biological Chemistry 03/2013;
• Article: Decorin potentiates IFN-γ activity in a model of allergic inflammation.

  Bocian C, Urbanowitz AK, Owens RT, Iozzo RV, Götte M, Seidler DG
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  ABSTRACT: The proteoglycan decorin modulates leukocyte recruitment during delayed-type hypersensitivity (DTH) responses. Decorin-deficient (Dcn-/-) mice show reduced edema formation during the first 24 h with a concurrent attenuated recruitment of Cd8+ leukocytes in the inflamed Dcn-/- ears. The aim of this study was to elucidate the molecular pathways affected by the loss of decorin. In vivo, reduced numbers of Cd8+ cells in Dcn-/- ears correlated with a reduced interferon-γ (Ifn-γ) expression. In vitro, Dcn-/- lymphocytes displayed an increased adhesion to brain microvascular (bEnd.3) endothelial cells. Decorin treatment of bEnd.3 increased Icam-I and down-regulated Vcam-I expression after Tnf-α stimulation. However, Dcn-/- and wild-type lymphocytes produced Ifn-γ after activation with Cd3ε. Upon incubation with decorin, endothelial cells and fibroblasts responded differently to Ifn-γ and Tnf-α: Ccl2 in bEnd.3 cells was more prominently up-regulated by Tnf-α compared to Ifn-γ. Notably, both factors were more potent in the presence of decorin. Compared to Tnf-α, Ifn-γ treatment induced significantly more CXCL-10, and both factors increased synthesis of CXCL-10 in the presence of decorin. The response to Ifn-γ was similar in Dcn-/- and wild-type fibroblasts, an additional source of CXCL-10. However, addition of decorin yielded significantly more CXCL-10. Notably, decorin increased the stability of Ifn-γ in vitro, and potentiated Ifn-γ-induced activation of Stat-1. Furthermore, only dermatan sulfate influenced Ifn-γ signaling by significantly increasing CXCL-10 expression in contrast to decorin protein core alone. Our data demonstrate that decorin modulates DTH responses by augmenting the induction of downstream effector cytokines of Ifn-γ and Tnf-α, thereby influencing the recruitment of Cd8+ lymphocytes into the inflamed tissue.
  Journal of Biological Chemistry 03/2013;
• Article: Regulatory role of proteasome in determination of platelet life span

  Manasa K. Nayak, Paresh P. Kulkarni, Debabrata Dash
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  ABSTRACT: Limit of platelet life span (8–10 days) is determined by the activity of a putative “internal clock” composed of Bcl-2 family proteins, whereas the role of other molecular players in this process remains obscure. Here, we sought to establish a central role of proteasome in platelet life span regulation. Administration of mice with inhibitors of proteasome peptidase activity induced significant thrombocytopenia. This was associated with enhanced clearance of biotin-labeled platelets from circulation and reduction in average platelet half-life from 66 to 37 h. Cells pretreated in vitro with proteasome inhibitors exhibited augmented annexin V binding and a drop in mitochondrial transmembrane potential indicative of apoptotic cell death and decreased platelet life span. These cells were preferentially phagocytosed by monocyte-derived macrophages, thus linking proteasome activity with platelet survival. The decisive role of proteasome in this process was underscored from enhanced expression of conformationally active Bax in platelets with attenuated proteasome activity, which was consistent with pro-apoptotic phenotype of these cells. The present study establishes a critical role of proteasome in delimiting platelet life span ostensibly through constitutive elimination of the conformationally active Bax. These findings bear potential implications in clinical settings where proteasome peptidase activities are therapeutically targeted.
  Journal of Biological Chemistry 03/2013; 288(10):6826-6834.
• Article: Immobilization of heparan sulfate on electrospun meshes to support embryonic stem cell culture and differentiation. Meade KA, White KJ, Pickford CE, Holley RJ, Marson A, Tillotson D, van Kuppevelt TH, Whittle JD, Day AJ, Merry CL. J Biol Chem. 2013 Feb 22;288(8):5530-8. doi: 10.1074/jbc.M112.423012. Epub 2012 Dec 12.

  Meade KA, White KJ, Pickford CE, Holley RJ, Marson A, Tillotson D, van Kuppevelt TH, Whittle JD, Day AJ, Merry CL
  Journal of Biological Chemistry 02/2013;
• Article: A novel distal enhancer confers chorionic expression on the human renin gene. J Biol Chem 1998 ; 273:25292-300.

  S Gernain, F Bonnet, S Fuchs, P Corvol, F Pinet
  Journal of Biological Chemistry 02/2013;
• Article: Endothelial Cell-Specific Chemotaxis Receptor (ECSCR) enhances Vascular Endothelial Growth Factor (VEGF) Receptor-2/Kinase insert domain receptor (KDR) activation and promotes proteolysis of internalized KDR

  Sreenivasulu Kilari, Indulekha Remadevi, Baofeng Zhao, Jing Pan, Robert Miao, Ramani Ramchandran
  Journal of Biological Chemistry 02/2013;
• Article: A Cdc42 GTPase Specific Inhibitor as a Molecular Probe and Prototype for Drug Development

  L. 2. Hong, S.R. Kenney, G.K. Philip, D. Simpson, C.E. Schroeder, J. Nöth, E. Romero, T. Buranda, A. Waller, J.J. Strouse, M. Carter, A. Chigaev, O. Ursu, T. Oprea, J.E. Golden, J. Aubé, L.G. Hudson, L.A. Sklar, A Wandinger-Ness
  Journal of Biological Chemistry 01/2013;