title: Mycobacterium tuberculosis AtsG (Rv0296c), GlmU (Rv1018c) and SahH (Rv3248c) Proteins Function as the Human IL-8-Binding Effectors and Contribute to Pathogen Entry into Human Neutrophils.
abstract: Mycobacterium tuberculosis is an extremely successful intracellular pathogen that has evolved a broad spectrum of pathogenic mechanisms that enable its manipulation of host defense elements and its survival in the hostile environment inside phagocytes. Cellular influx into the site of mycobacterial entry is mediated by a variety of chemokines, including interleukin-8 (IL-8), and the innate cytokine network is critical for the development of an adaptive immune response and infection control. Using affinity chromatography, liquid chromatography electrospray ionization tandem mass spectrometry and surface plasmon resonance techniques, we identified M. tuberculosis AtsG arylsulphatase, bifunctional glucosamine-1-phosphate acetyltransferase and N-acetylglucosamine-1-phosphate uridyl transferase (GlmU) and S-adenosyl-L-homocysteine hydrolase (SahH) as the pathogen proteins that bind to human IL-8. The interactions of all of the identified proteins (AtsG, GlmU and SahH) with IL-8 were characterized by high binding affinity with KD values of 6.83x10-6 M, 5.24x10-6 M and 7.14x10-10 M, respectively. Furthermore, the construction of Mtb mutant strains overproducing AtsG, GlmU or SahH allowed determination of the contribution of these proteins to mycobacterial entry into human neutrophils. The significantly increased number of intracellularly located bacilli of the overproducing M. tuberculosis mutant strains compared with those of "wild-type" M. tuberculosis and the binding interaction of AtsG, GlmU and SahH proteins with human IL-8 may indicate that these proteins participate in the modulation of the early events of infection with tubercle bacilli and could affect pathogen attachment to target cells.
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
authors.forename: Bozena
authors.initials: B
authors.lastname: Dziadek
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Brzostek
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Grzybowski
authors.affiliation: Department of Immunology and Infectious Biology, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
authors.forename: Marek
authors.initials: M
authors.lastname: Fol
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Krupa
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland.
authors.forename: Jakub
authors.initials: J
authors.lastname: Kryczka
authors.affiliation: Laboratory of RNA Biology and Functional Genomics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Plocinski
authors.affiliation: Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United States of America.
authors.forename: Anna
authors.initials: A
authors.lastname: Kurdowska
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland.
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Dziadek
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D016209:Interleukin-8 / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D009504:Neutrophils / Q000382:microbiology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014376:Tuberculosis / Q000276:immunology / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D007106:Immune Sera; D016209:Interleukin-8; D011994:Recombinant Proteins
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