DOCSLIB.ORG

Ο-Sialoglycoprotein Endopeptidase

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Ο-Sialoglycoprotein Endopeptidase Ο-Sialoglycoprotein Endopeptidase Ο-sialoglycoprotein endopeptidase is a neutral metalloprotease purified from Mannheimia haemolytica (formerly known as Pasteurella haemolytica).This unique proteolytic enzyme specifically cleaves proteins bearing clusters of negatively charged sugars: Ο-sialoglycoproteins and sulfated glycoproteins. The enzyme is inhibited at high concentrations of EDTA (above 10 mM), by sialate analogues (above 5 mM) and by the putative metal ion activator Zn2+ at concentrations around 50 μM. No direct activation by zinc ions has been observed at any concentration. The enzyme is not affected by serine protease inhibitors, aspartyl protease inhibitors or thiol inhibitors. Numerous Ο-sialoglycoproteins have been shown to be cleaved by this enzyme, but no N-linked sialoglycoproteins or unglycosylated proteins have been found to be substrates. The list of Ο-sialoglycoprotein endopeptidase substrates includes: human RBC glycophorin A the human antigens CD34, CD43, CD44, CD45 the IL-7 receptor the receptors for E-selectin, L-selectin and P-selectin the human tumor antigens epiglycanin and epitectin the platelet glycoprotein 1ba Ο-sialoglycoprotein the laminin binding proteins, dystroglycan and cranin Product endopeptidase, the lymphocyte adhesion molecule VAP-1 Lyophilized viral glycoproteins; and bone sialoprotein the sulfated glycoprotein CD24 Size 1.2 mg Applications: Cat. # CLE100 Ideal for characterizing cell surface glycoproteins. Useful in glycoprotein epitope-mapping studies. Can be used to modify the adhesion properties of cells, including rolling behaviour of neutrophils. Can be used to degrade Ο-sialoglycoproteins to enable peptide sequencing of the resultant fragments. Used for the immunomagnetic separation of human ISO 9001:2008 and ISO 13485:2003 registered. stem cells bearing the CD34 antigen, in that it will cleave Toll Free: CD34 and release the antibody-magnetic bead complex from the isolated stem cell. In Canada: 1-800-268-5058 In the U.S.A.: 1-800-721-1644 O-Sialoglycoprotein Endopeptidase Catalogue # CLE100, O-sialoglycoprotein endopeptidase, Lyophilized, 1.2 mg, $398 O-sialoglycoprotein endopeptidase is a neutral metalloprotease purified from Mannheimia haemolytica (formerly known as Pasteurella haemolytica). This unique proteolytic enzyme specifically cleaves proteins bearing clusters of negatively charged sugars, so that O-sialoglycoproteins and sulfated glycoproteins are good substrates. The enzyme is inhibited at high concentrations of EDTA (above 10 mM), by sialate analogues (above 5mM) and the putative metal ion activator Zn2+ at concentrations around 50 M. No direct activation by zinc ions has been observed at any concentration. The enzyme is not affected by serine protease inhibitors, aspartyl protease inhibitors or thiol inhibitors. Numerous O-sialoglycoproteins have been shown to be cleaved by this enzyme, but no N-linked sialoglycoproteins or unglycosylated proteins have been found to be substrates. The list of O-sialoglycoprotein substrates includes human RBC glycophorin A; the human antigens CD34, CD43, CD44, CD45; the IL-7 receptor; the receptors for E- selectin, L-selectin and P-selectin; the human tumor antigens epiglycanin and epitectin; the platelet glycoprotein 1ba; the laminin binding proteins, dystroglycan and cranin; the lymphocyte adhesion molecule VAP-1; viral glycoproteins; and bone sialoprotein. A sulfated glycoprotein, CD24, is also cleaved by the enzyme. Applications: - Ideal for characterizing cell surface glycoproteins. - Useful in glycoprotein epitope-mapping studies. - Can be used to modify the adhesion properties of cells, including rolling behavior of neutrophils. - Can be used to degrade O-sialoglycoproteins to enable peptide sequencing of the resultant fragments. - Used for the immunomagnetic separation of human stem cells bearing the CD34 antigen, in that it will cleave CD34 and release the antibody-magnetic bead complex from the isolated stem cell. GlycoproteolyticGlycoproteolytic Activity against Various Activity Substrates: Against Various Substrates: 1800 0.6 1500 1200 mol/liter/min) μ 0.4 Glycophorin A 0.008 900 0.006 0.004 1/F Fluorescence 600 0.2 0.002 κ–Casein 0.000 300 0 2 4 6 [1/S] μg -1 Substrate degraded ( degraded Substrate 0.00 0 5 10 15 2 4 6 8 10 125 FigureFigure 1. A) Hydrolysis A Substrate of human (glycophorinμg) A comparedFigure with that B of bovine [BODIPY-FL-GPA]K-casein: Hydrolysis μg of the two I- labeled substrates as a function of substrate concentration in a 5 min incubation with equivalent amounts of P. haemolytica glycoprotease pH 4.5 precipitate. Hydrolysis was measured by the disappearance of TCA-soluble productsFigure A.from Hydrolysis 125I-labeled of human glycophorin glycophorin AA comparedbands from with thatSDS of-PAGE bovine gels K-casein: and Hydrolysisby the ofappearance the two 125I-labeled of TCA substrates-soluble as a function of substrate125 concentration in[20] a 5 min incubation with equivalent amounts of P. haemolytica glycoprotease pH 4.5 precipitate. productsHydrolysis from was measuredI-labeled by -thecasein. disappearance B) Substrate of TCA-soluble concentration products from dependence 125I-labeled forglycophorin the glycoprotease A bands from hydrolysis SDS-PAGE gels of and by the appearance of TCA-soluble products from 125I-labeled κ-casein.[20] Figure B. Substrate concentration dependence for the glycoprotease hydrolysis of BODIPY-FL–GPA. The 25 μl reaction mixture containing 0.4 μg concentrated culture supernatant glycoprotease, incubated with varying amounts of BODIPY-FL–GPA in 50 mM HEPES buffer at 37°C for 10 min. The reaction was stopped by dilution and the fluorescence was measured. The inset shows the double reciprocal plot for the same data. Means + SE (n = 3).[19] References: 1. G.L. Otulakowski, P.E. Shewen, A.E. Udoh, A. Mellors and B.N. Wilkie (1983). 10. J.G. Oriol, K.J. Betteridge, A.J. Clarke and F.J. Sharom (1993). Mucin-like glycoproteins in the equine Proteolysis of sialo-glycoprotein by Pasteurella haemolytica cytotoxic culture supernatants. embryonic capsule. Mol Reprod. Develop. 34, 255-265. Infection and Immunity, 42, 64-70. 11. J.C.W. Marsh, D.R. Sutherland, J. Davidson, A. Mellors & A. Keating (1992). Retention of progenitor cell 2. K.M. Abdullah, R.Y.C. Lo, & A. Mellors (1991). Cloning, nucleotide sequence and expression of the function in CD34+ cells purified using a novelΟ -sialoglycoprotease. Leukemia 6, 926-934. P. haemolytica A1 glycoprotease gene. J. Bacteriol. 173, 5597-5603. 12. D.R. Sutherland, J.C.W. Marsh, J. Davidson, M.A. Baker, A. Keating, and A. Mellors (1992). 3. K.M. Abdullah, E.A. Udoh, P.E. Shewen, and A. Mellors (1991). A neutral glycoprotease of Differential Sensitivity of CD34 Epitopes to Cleavage by Pasteurella haemolytica Glycoprotease: P. haemolytica A1 specifically cleavesΟ -sialoglycoproteins. Infect. Immun. 60, 56-62. Implications for Purification of CD34-positive Progenitor Cells. Exp. Hematol. 20, 590-599. 4. D.R. Sutherland, K.M. Abdullah, P. Cyopick, & A. Mellors (1992). Cleavage of the cell-surface 13. Sutherland D.R. et al. (1996). Identification of CD34+ subsets after glycoprotease selection: Ο-sialoglycoproteins CD34, CD43, CD44, CD45 by a novel glycoprotease from P. haemolytica. Engraftment of CD34+ Thy-1+Lin- stem cells in fetal sheep. Exp. Hematol. 24, 795-806. J. Immunol. 148, 1458-1464. 14. A. Mellors and D.R. Sutherland (1994). Tools to cleave glycoproteins. Trends in Biotechnol. 12, 15-18. 5. C.N. Steininger, C.A. Eddy, R.M. Leimgruber, A. Mellors and J.K. Welply (1992). The glycoprotease of 15. W. M. Cladman, M. –A.V. Watt, J. –P. Dini, and A. Mellors (1996). The Pasteurella haemolytica P. haemolytica A1 eliminates binding of myeloid cells to P-selectin but not to E-selectin. Ο-sialoglycoprotein endopeptidase is inhibited by zinc ions and does not cleave fetuin. Biochem. Biophys. Res. Commun. 188, 760-766. Biochem. Biophys. Res. Commun. 220, 141-146. 6. H. Nakada, M. Inouue, Y. Numata, N. Tanaka, I. Funakoshi, S. Fukui, A. Mellors and I. Yamashina (1993) 16. M. –A.V. Watt, R.Y.C. Lo, and A. Mellors (1997). Refolding of recombinant Pasteurella haemolytica Epitope structure of Tn glycophorin A for anti-Tn antibody (MLS 128). Proc. Natl. Acad. A1 glycoprotease expressed in an E. coli thioredoxin gene fusion system. Cell Stress and Sci. USA 90, 2495-2499. (This paper shows how the enzyme can be used to identify specific Chaperones 2, 180-190. epitope domains on human erythrocyte glycophorin). 17. P. Jiang and A. Mellors (2004). Ο-Sialoglycoprotein endopeptidase. In Handbook of Proteolytic 7. L. Healy and I. Titley (1993). The P. haemolytica glycoprotease cleaves the Interleukin 7 receptor Enzymes: Second Edition ed. A.L. Barrett and J.F. Woessner, p.977-980, Elsevier, London. expressed on human lymphoid cells. Personal communication, with permission to quote the findings. 18. M. –A.V. Watt, A. Mellors and R.Y.C. Lo (1997). Comparison of the recombinant and authentic forms 8. R.H. Hu, A. Mellors and V.P. Bhavanandan (1992). Selective cleavage of epitectin from the surface of the P. haemolytica A1 glycoprotease. FEMS Microbiology Lett. 47, 37-43. of H.Ep.2 human laryngeal carcinoma cells by a glycoprotease from P. haemolytica. 19. P. Jiang and A. Mellors (1998). Membrane protein proteolysis assayed by fluorescence quenching: Glycobiology 2, 500. assay of Ο-sialoglycoprotein endopeptidase. Anal. Biochem. 259, 8-15. Figure B is reprinted for the 9. E. Yeo, A. Mellors and R. Sutherland (1993). Cleavage of leukocyte surface
Load more

Recommended publications
  • Soluble Factor in Normal Tissues That Stimulates High-Molecular-Weight Sialoglycoprotein Production by Human Colon Carcinoma Cells1
    (CANCER RESEARCH 50, 3331-3338. June I, I990| Soluble Factor in Normal Tissues That Stimulates High-Molecular-Weight Sialoglycoprotein Production by Human Colon Carcinoma Cells1 Tatsuro Irimura,2 Andrew M. Mclsaac, Debora A. Carlson, Masato Vagita, Elizabeth A. Grimm, David G. Menter, David M. Ota, and Karen R. Cleary Departments of Tumor Biology IT. I., A. M., D. A. C., M. Y., E. A. G., D. G. M.], General Surgery /E. A. G., D. M. O.J, and Pathology [K. R. C.J, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030 ABSTRACT linked carbohydrate chains. Secreted mucins are believed to function as protective molecules and as lubricants on tissue The stimulation of high molecular »eight Sialoglycoprotein synthesis surfaces. Despite previous attempts by several laboratories to by a soluble factor derived from normal colon tissues was studied in vitro with human colon carcinoma cell lines, HT-29 P and a metastatic variant classify and characterize colorectal mucins, it remained unclear HT-29 LMM. The synthesis of all three high-molecular-weight sialo- whether a specific biological function is associated with unique glycoproteins (approximate M, 900,000, 740,000, and 450,000) by HT- carbohydrate structures in these mucins (10,13-22). Our recent 29 P cells or HT-29 LMM cells growing in vitro was enhanced by studies utilizing pathological specimens of colorectal carcinoma supplementing the culture medium with a conditioned medium of fresh suggested that at least four mucin-like glycoproteins with dif human colon organ culture. Changes were detected by polyacrylamide ferent carbohydrate chains were independently regulated and gel electrophoresis of lysates from [3H)glucosamine-labeled cells on 3% either directly or inversely correlated with the metastatic poten gels followed by fluorography, or by electrophoresis of lysates from tial of these malignant tumors (23-27).3 Thus, these changes unlabeled cells followed by incubation with '"I-labeled wheat germ were apparently associated with the progression of colon car agglutinin and autoradiography.
    [Show full text]
  • Sialylated Keratan Sulfate Chains Are Ligands for Siglec-8 in Human Airways
    Sialylated Keratan Sulfate Chains are Ligands for Siglec-8 in Human Airways by Ryan Porell A dissertation submitted to Johns Hopkins University in conformity with the requirements for the degree of Doctor of Philosophy Baltimore, Maryland September 2018 © 2018 Ryan Porell All Rights Reserved ABSTRACT Airway inflammatory diseases are characterized by infiltration of immune cells, which are tightly regulated to limit inflammatory damage. Most members of the Siglec family of sialoglycan binding proteins are expressed on the surfaces of immune cells and are immune inhibitory when they bind their sialoglycan ligands. When Siglec-8 on activated eosinophils and mast cells binds to its sialoglycan ligands, apoptosis or inhibition of mediator release is induced. We identified human airway Siglec-8 ligands as sialylated and 6’-sulfated keratan sulfate (KS) chains carried on large proteoglycans. Siglec-8- binding proteoglycans from human airways increase eosinophil apoptosis in vitro. Given the structural complexity of intact proteoglycans, target KS chains were isolated from airway tissue and lavage. Biological samples were extensively proteolyzed, the remaining sulfated glycan chains captured and resolved by anion exchange chromatography, methanol-precipitated then chondroitin and heparan sulfates enzymatically hydrolyzed. The resulting preparation consisted of KS chains attached to a single amino acid or a short peptide. Purified KS chains were hydrolyzed with either hydrochloric acid or trifluoroacetic acid to release acidic and neutral sugars, respectively, followed by DIONEX carbohydrate analysis. To isolate Siglec-8-binding KS chains, purified KS chains from biological samples were biotinylated at the amino acid, resolved by affinity and/or size- exclusion chromatography, the resulting fractions immobilized on streptavidin microwell plates, and probed for binding of Siglec-8-Fc.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 7,998,740 B2 Sackstein (45) Date of Patent: Aug
    US007998.740B2 (12) United States Patent (10) Patent No.: US 7,998,740 B2 Sackstein (45) Date of Patent: Aug. 16, 2011 (54) CYTOKINE INDUCTION OF SELECTIN Butcher, E. C., "Leukocyte-endothelial cell recognition: three (or LGANDS ON CELLS more) steps to specificity and diversity”. Cell, 67: 1033-6 (1991). Cowlandet al., “Isolation of neutrophil precursors from bone marrow (76) Inventor: Robert Sackstein, Sudbury, MA (US) for biochemical and transcriptional analysis”. J. Immunol. Meth. 232:191-200 (1999). (*) Notice: Subject to any disclaimer, the term of this Dereure et al., “Neutrophil-dependent cutaneous side-effects of patent is extended or adjusted under 35 leucocyte colony-stimulating factors: manifestations of a neutrophil U.S.C. 154(b) by 325 days. recovery syndrome'. Brit. J. Dermatol., 150: 1228-30 (2004). Dimitroff et al., “CD44 is a major E-selectin ligand on human (21) Appl. No.: 11/779,650 hematopoietic progenitor cells'. J. Cell. Biol. 153:1277-86 (2001). Dimitroffet al., “A distinct glycoform of CD44 is an L-selectin ligand (22) Filed: Jul.18, 2007 on human hematopoietic cells'. Proc. Natl. Acad. Sci. USA, (65) Prior Publication Data 97:13841-6 (2000). Dimitroffet al., “Differential L-selectin binding activities of human US 2009/00531.98 A1 Feb. 26, 2009 hematopoietic cell L-selectin ligands, Hcell and PSGL-1”. J. Biol. Chem., 276:47623-31 (2001). Related U.S. Application Data Elfenbein et al., “Primed marrow for autologous and allogeneic trans (60) Provisional application No. 60/831,525, filed on Jul. plantation: a review comparing primed marrow to mobilized blood 18, 2006. and steady-state marrow”.
    [Show full text]
  • Adherence of Mycoplasma Gallisepticum to Human Erythrocytes M
    INFECTION AND IMMUNITY, Aug. 1978, p. 365-372 Vol. 21, No. 2 0019-9567/78/0021-0365$02.00/0 Copyright i 1978 American Society for Microbiology Printed in U.S.A. Adherence of Mycoplasma gallisepticum to Human Erythrocytes M. BANAI,1 I. KAHANE,' S. RAZINl* AND W. BREDT2 Biomembrane Research Laboratory, Department of Clinical Microbiology, The Hebrew University- Hadassah Medical School, Jerusalem, Israel,' and Institute for General Hygiene and Bacteriology, Center for Hygiene, Albert-Ludwigs- Universitat, D- 7800 Freiburg, West Germany Received for publication 7 February 1978 Pathogenic mycoplasmas adhere to and colonize the epithelial lining of the respiratory and genital tracts ofinfected animals. An experimental system suitable for the quantitative study of mycoplasma adherence has been developed by us. The system consists of human erythrocytes (RBC) and the avian pathogen Mycoplasma gallisepticum, in which membrane lipids were labeled. The amount of mycoplasma cells attached to the RBC, which was determined according to radioactivity measurements, decreased on increasing the pH or ionic strength of the attachment mixture. Attachment followed first-order kinetics and depended on temperature. The mycoplasma cell population remaining in the supernatant fluid after exposure to RBC showed a much poorer ability to attach to RBC during a second attachment test, indicating an unequal distribution of binding sites among cells within a given population. The gradual removal of sialic acid residues from the RBC by neuraminidase was accompanied by a decrease in mycoplasma attachment. Isolated glycophorin, the RBC membrane glycoprotein carrying almost all the sialic acid moieties ofthe RBC, inhibited M. gallisepticum attachment, whereas asialoglycophorin and sialic acid itself were very poor inhibitors of attachment.
    [Show full text]
  • General Considerations of Coagulation Proteins
    ANNALS OF CLINICAL AND LABORATORY SCIENCE, Vol. 8, No. 2 Copyright © 1978, Institute for Clinical Science General Considerations of Coagulation Proteins DAVID GREEN, M.D., Ph.D.* Atherosclerosis Program, Rehabilitation Institute of Chicago, Section of Hematology, Department of Medicine, and Northwestern University Medical School, Chicago, IL 60611. ABSTRACT The coagulation system is part of the continuum of host response to injury and is thus intimately involved with the kinin, complement and fibrinolytic systems. In fact, as these multiple interrelationships have un­ folded, it has become difficult to define components as belonging to just one system. With this limitation in mind, an attempt has been made to present the biochemistry and physiology of those factors which appear to have a dominant role in the coagulation system. Coagulation proteins in general are single chain glycoprotein molecules. The reactions which lead to their activation are usually dependent on the presence of an appropriate surface, which often is a phospholipid micelle. Large molecular weight cofactors are bound to the surface, frequently by calcium, and act to induce a favorable conformational change in the reacting molecules. These mole­ cules are typically serine proteases which remove small peptides from the clotting factors, converting the single chain species to two chain molecules with active site exposed. The sequence of activation is defined by the enzymes and substrates involved and eventuates in fibrin formation. Mul­ tiple alternative pathways and control mechanisms exist throughout the normal sequence to limit coagulation to the area of injury and to prevent interference with the systemic circulation. Introduction RatnofP4 eloquently indicates in an arti­ cle aptly entitled: “A Tangled Web.
    [Show full text]
  • Mucin-Selective Protease Stce Enables Molecular and Functional Analysis of Human Cancer-Associated Mucins
    The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins Stacy A. Malakera,1, Kayvon Pedrama,1, Michael J. Ferracaneb, Barbara A. Bensingc, Venkatesh Krishnand, Christian Pette,f, Jin Yue, Elliot C. Woodsa, Jessica R. Kramerg, Ulrika Westerlinde,f, Oliver Dorigod, and Carolyn R. Bertozzia,h,2 aDepartment of Chemistry, Stanford University, Stanford, CA 94305; bDepartment of Chemistry, University of Redlands, Redlands, CA 92373; cDepartment of Medicine, San Francisco Veterans Affairs Medical Center and University of California, San Francisco, CA 94143; dStanford Women’s Cancer Center, Division of Gynecologic Oncology, Stanford University, Stanford, CA 94305; eLeibniz-Institut für Analytische Wissenschaften (ISAS), 44227 Dortmund, Germany; fDepartment of Chemistry, Umeå University, 901 87 Umeå, Sweden; gDepartment of Bioengineering, University of Utah, Salt Lake City, UT 84112; and hHoward Hughes Medical Institute, Stanford, CA 94305 Edited by Laura L. Kiessling, Massachusetts Institute of Technology, Cambridge, MA, and approved February 25, 2019 (received for review July 30, 2018) Mucin domains are densely O-glycosylated modular protein domains When strung together in “tandem repeats” mucin domains can that are found in a wide variety of cell surface and secreted proteins. form the large structures characteristic of mucin family proteins. Mucin-domain glycoproteins are known to be key players in a host Mucins can be hundreds to thousands of amino acids long of human diseases, especially cancer, wherein mucin expression and and >50% glycosylation by mass (11); MUC16, one of the largest glycosylation patterns are altered. Mucin biology has been difficult mucins, can exceed 22,000 residues and 85% glycosylation by mass to study at the molecular level, in part, because methods to manip- (12), with a persistence length of 1–5 μm (13).
    [Show full text]
  • Review Article B Lymphocytes: Development, Tolerance, and Their Role in Autoimmunity—Focus on Systemic Lupus Erythematosus
    Hindawi Publishing Corporation Autoimmune Diseases Volume 2013, Article ID 827254, 17 pages http://dx.doi.org/10.1155/2013/827254 Review Article B Lymphocytes: Development, Tolerance, and Their Role in Autoimmunity—Focus on Systemic Lupus Erythematosus Gabriel J. Tobón, Jorge H. Izquierdo, and Carlos A. Cañas Department of Internal Medicine, Division of Rheumatology, Fundacion´ Valle del Lili, ICESI University School of Medicine, Cra 98 No. 18-49, Cali, Colombia Correspondence should be addressed to Gabriel J. Tobon;´ [email protected] Received 30 June 2013; Accepted 6 August 2013 Academic Editor: Juan-Manuel Anaya Copyright © 2013 Gabriel J. Tobon´ et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. B lymphocytes are the effectors of humoral immunity, providing defense against pathogens through different functions including antibody production. B cells constitute approximately 15% of peripheral blood leukocytes and arise from hemopoietic stem cells in the bone marrow. It is here that their antigen receptors (surface immunoglobulin) are assembled. In the context of autoimmune diseases defined by B and/or T cell autoreactive that upon activation lead to chronic tissue inflammation and often irreversible structural and functional damage, B lymphocytes play an essential role by not only producing autoantibodies but also functioning as antigen-presenting cells (APC) and as a source of cytokines. In this paper, we describe B lymphocyte functions in autoimmunity and autoimmune diseases with a special focus on their abnormalities in systemic lupus erythematosus. 1. Introduction memory B cells.
    [Show full text]
  • The Membrane-Bound Mucins
    The Membrane-Bound Mucins: How Large O-Glycoproteins Play Key Roles in Epithelial Cancers and Hold Promise as Biological Tools for Gene-Based and Immunotherapies Nicolas Jonckheere, Isabelle Seuningen To cite this version: Nicolas Jonckheere, Isabelle Seuningen. The Membrane-Bound Mucins: How Large O-Glycoproteins Play Key Roles in Epithelial Cancers and Hold Promise as Biological Tools for Gene-Based and Immunotherapies. Critical Review in Oncogenesis, Begell House, 2008, 14 (2-3), pp.177-196. 10.1615/CritRevOncog.v14.i2-3.30. hal-02905788 HAL Id: hal-02905788 https://hal.archives-ouvertes.fr/hal-02905788 Submitted on 29 Sep 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Critical Reviews™ in Oncogenesis, 14(2&3), x–xx (2008) [[AU: PLEASE NOTE HIGHLIGHTED REVISIONS FOR YOUR REVIEW. THANK YOU.]] The Membrane-Bound Mucins: How Large O-Glycoproteins Play Key Roles in Epithelial Cancers and Hold Promise as Biological Tools for Gene and Immunotherapies Nicolas Jonckheere & Isabelle Van Seuningen Inserm, U837, Centre de Recherche Jean-Pierre Aubert, Place de Verdun, Lille, F-59045 France Address all correspondence to Dr. Isabelle Van Seuningen, Inserm U837, Team 5 “Mucins, epithelial dif- ferentiation and carcinogenesis,” Centre de Recherche Jean-Pierre Aubert, Place de Verdun, 59045 Lille cedex, France; [email protected] ABSTRACT: Membrane-bound mucins belong to an ever-increasing family of O-glycopro- teins that share a structure conserved throughout evolution.
    [Show full text]
  • Transforming the Cell Surface Through Proteolysis Laura L Kiessling and Eva J Gordon
    Review R49 Transforming the cell surface through proteolysis Laura L Kiessling and Eva J Gordon Protein shedding, or the proteolytic cleavage of a protein from Introduction the surface of a cell, is emerging as an important mechanism in Cells have the remarkable ability to direct the functions the regulation of cellular activity but it is poorly understood. and activities of their thousands of proteins through Growing evidence suggesting that protein shedding and several mechanisms. One of these is phosphorylation - a protein function are closely linked may lead to new strategies reversible modification that affects function by changing for the treatment of a wide range of diseases. protein-protein interaction propensities. An alternative and more permanent way to direct cellular processes is by Address: Departments of Chemistry and Biochemistry, University of proteolysis. Intracellularly, proteolysis can regulate the Wisconsin-Madison, Madison, WI 53706, USA. presence of specific proteins, influencing fundamental Correspondence: Laura L Kiessling processes such as the cell cycle or apoptosis, and proteoly- E-mail: [email protected] sis regulates important processes outside the cell, such as Chemistry 81 Biology March 1998,5:R49-R62 cell migration and wound repair. Proteolysis can also func- http://biomednet.com/elecref/10745521005R0049 tion as a control mechanism on the cell surface. For example, the G-protein-coupled thrombin receptor can be 0 Current BioioQy Ltd ISSN 1074-5521 activated by limited proteolysis [l]: thrombin cuts the amino terminus of its receptor, causing a conformational change that activates the receptor. Thus, regulated prote- olysis has an important role on the cell surface. This review focuses on the importance of proteolysis in the release of the extracellular portion of a protein by cleavage at the cell surface, referred to here as protein shedding.
    [Show full text]
  • Siglec-8 Ligands in Human Airway and Airway Secretions
    SIGLEC-8 LIGANDS IN HUMAN AIRWAY AND AIRWAY SECRETIONS by Anabel G. Alvarenga A dissertation submitted to Johns Hopkins University in conformity with the requirements for the degree of Doctor of Philosophy Baltimore, Maryland April, 2018 ABSTRACT Siglecs – sialic acid binding Ig-like lectins – are regulatory molecules expressed on subsets of immune cells where most inhibit inflammation when engaged by complementary sialoglycan ligands on target tissues. Both eosinophils and mast cells express Siglec-8 on the cell surface, and when Siglec-8 binds to sialoglycan ligands on tissues apoptosis of eosinophils and inhibition of mediator release by mast cells is induced, limiting inflammation. Although Siglec-8 has been shown to bind a synthetic glycan 6’-sulfated sialyl N-acetyllactosamine, the endogenous Siglec-8 ligand in human airways was still unknown. This study provides evidence of endogenous high molecular weight Siglec-8 ligands in human airway and airway secretions that are sensitive to sialidase and keratanase treatment. Siglec-8 ligands were isolated and identified from normal postmortem human airways or from nasal lavage. Glycoproteins were separated by size exclusion chromatography and resolved by composite agarose-acrylamide gel electrophoresis, blotted, and probed with human Fc-tagged Siglec-8, revealing three binding species (270 kDa, 600 kDa and 1000 kDa) in tracheal extracts and one major binding specie (~900 kDa) in nasal lavage. Ligand- containing fractions were pooled, and ligands were captured by immunoprecipitation using His-tagged pentameric Siglec-8 bound to nickel-Sepharose beads. Siglec-8-precipitated ligands were subjected to mass spectrometric proteomic analysis, revealing the proteoglycan aggrecan as the predominant protein in all three-size species of Siglec-8 ligands extracted from trachea and glycoprotein-340 as the predominant protein in sample purified from nasal lavage.
    [Show full text]
  • ADAM9) Required
    bioRxiv preprint doi: https://doi.org/10.1101/491068; this version posted December 9, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Identification of A Disintegrin and Metalloproteinase 9 domain (ADAM9) required 2 in the early stages of encephalomyocarditis virus infection 3 Running title: ADAM9 is a major encephalomyocarditis virus receptor 4 Lindsey E. Bazzonea*, Michael Kinga*, Christopher R. MacKaya*, Pyae P. Kyawea, Paul 5 Meranerb, Daniel Lindstroma, Joselyn Rojas-Quinteroc, Caroline A. Owenc, Jennifer P. 6 Wanga,d, Abraham L. Brassa,b, Evelyn A. Kurt-Jonesa,d,#, Robert W. Finberga,d,# 7 aDepartment of Medicine, University of Massachusetts Medical School, Worcester, MA 8 01605, USA 9 bDepartment of Microbiology and Physiological Systems (MaPS), University of 10 Massachusetts Medical School, Worcester, MA 01655, USA 11 cDivision of Pulmonary and Critical Care Medicine, Brigham and Women’s Hospital and 12 Harvard Medical School, Boston, MA 02115, USA 13 dProgram in Innate Immunity, University of Massachusetts Medical School, Worcester, 14 MA 01605, USA 15 16 17 18 *Contributed equally 19 #Co-corresponding authors: [email protected] and 20 [email protected] 21 Abstract word count: 165 words 22 Main text word count: 5,097 words bioRxiv preprint doi: https://doi.org/10.1101/491068; this version posted December 9, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 23 Abstract 24 Encephalomyocarditis virus (EMCV) is a picornavirus that produces lytic 25 infections in murine and human cells.
    [Show full text]
  • Gene Symbol Genbank MCF7-T MCF-F De
    Table S2. Genes with altered expression in MCF7-T and MCF7-F cells Fold Change (vs. MCF7) Gene symbol Genbank MCF7-T MCF-F Description Downregulated in both MCF7-T and MCF7-F ABAT NM_020686 -3.10 -4.55 4-aminobutyrate aminotransferase ASCL2 NM_005170 -3.58 -6.11 Achaete-scute complex-like 2 (Drosophila) AZGP1 NM_001185 -29.21 -5.09 Alpha-2-glycoprotein 1, zinc CAP2 NM_006366 -3.23 -6.03 CAP, adenylate cyclase-associated protein, 2 (yeast) CD24 NM_013230 -4.20 -3.89 CD24 antigen (small cell lung carcinoma cluster 4 antigen) CDC42EP5 NM_145057 -3.89 -6.01 CDC42 effector protein (Rho GTPase binding) 5 CISH NM_145071 -4.76 -3.03 Cytokine inducible SH2-containing protein CLDN3 NM_001306 -4.24 -3.05 Claudin 3 CRIP1 NM_001311 -3.70 -12.43 Cysteine-rich protein 1 (intestinal) CRIP2 NM_001312 -5.03 -5.30 Cysteine-rich protein 2 CTXN1 Hs.250879 -3.53 -3.21 Cortexin 1 DLC1 NM_182643 -4.03 -21.61 Deleted in liver cancer 1 DNAJC12 NM_201262 -3.17 -67.76 DnaJ (Hsp40) homolog, subfamily C, member 12 EFEMP1 NM_018894 -5.30 -4.99 EGF-containing fibulin-like extracellular matrix protein 1 EFS NM_032459 -3.31 -25.59 Embryonal Fyn-associated substrate FHL1 NM_001449 -24.97 -110.29 Four and a half LIM domains 1 FLJ23548 NM_024590 -8.42 -8.30 Arylsulfatase J FOLR1 NM_016731 -4.56 -4.54 Folate receptor 1 (adult) GREB1 NM_148903 -97.58 -59.66 GREB1 protein HEY2 NM_012259 -7.29 -8.42 Hairy/enhancer-of-split related with YRPW motif 2 HRASLS3 NM_007069 -3.51 -4.84 HRAS-like suppressor 3 INHBB NM_002193 -5.74 -3.73 Inhibin, beta B (activin AB beta polypeptide) KLK10
    [Show full text]