Proteomic profiling of cell envelope-associated proteins fromStaphylococcus aureus

PROTEOMICS ◽  
2006 ◽  
Vol 6 (5) ◽  
pp. 1530-1549 ◽  
Author(s):  
Christine L. Gatlin ◽  
Rembert Pieper ◽  
Shih-Ting Huang ◽  
Emmanuel Mongodin ◽  
Elizabeth Gebregeorgis ◽  
...  
Keyword(s):  
Cell Envelope ◽  
Proteomic Profiling ◽  
Associated Proteins

scholarly journals Compartmentalized Cell Envelope Biosynthesis in Mycobacterium tuberculosis

2022 ◽  
Author(s):  
Julia Puffal ◽  
Ian L. Sparks ◽  
James R. Brenner ◽  
Xuni Li ◽  
John D. Leszyk ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Proteomic Analysis ◽  
Fluorescent Protein ◽  
Cell Envelope ◽  
Membrane Domain ◽  
Protein Fusion ◽  
Final Maturation ◽  
Associated Proteins ◽  
Gradient Fractionation ◽  
Membrane Compartmentalization

The intracellular membrane domain (IMD) is a metabolically active and laterally discrete membrane domain initially discovered in Mycobacterium smegmatis. The IMD correlates both temporally and spatially with the polar cell envelope elongation in M. smegmatis. Whether or not a similar membrane domain exists in pathogenic species remains unknown. Here we show that the IMD is a conserved membrane structure found in Mycobacterium tuberculosis. We used two independent approaches, density gradient fractionation of membrane domains and visualization of IMD-associated proteins through fluorescence microscopy, to determine the characteristics of the plasma membrane compartmentalization in M. tuberculosis. Proteomic analysis revealed that the IMD is enriched in metabolic enzymes that are involved in the synthesis of conserved cell envelope components such as peptidoglycan, arabinogalactan, and phosphatidylinositol mannosides. Using a fluorescent protein fusion of IMD-associated proteins, we demonstrated that this domain is concentrated in the polar region of the rod-shaped cells, where active cell envelope biosynthesis is taking place. Proteomic analysis further revealed the enrichment of enzymes involved in synthesis of phthiocerol dimycocerosates and phenolic glycolipids in the IMD. We validated the IMD association of two enzymes, α1,3-fucosyltransferase and fucosyl 4-O-methyltransferase, which are involved in the final maturation steps of phenolic glycolipid biosynthesis. Taken together, these data indicate that functional compartmentalization of membrane is an evolutionarily conserved feature found in both M. tuberculosis and M. smegmatis, and M. tuberculosis utilizes this membrane location for the synthesis of its surface-exposed lipid virulence factors.

scholarly journals Longitudinal proteomic profiling reveals increased early inflammation and sustained apoptosis proteins in severe COVID-19

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Liis Haljasmägi ◽  
Ahto Salumets ◽  
Anna Pauliina Rumm ◽  
Meeri Jürgenson ◽  
Ekaterina Krassohhina ◽  
...  
Keyword(s):  
Plasma Proteins ◽  
Proteomic Profiling ◽  
Apoptotic Pathway ◽  
Neutralization Capacity ◽  
Inflammatory Conditions ◽  
Associated Proteins ◽  
Life Threatening ◽  
Inflammatory Milieu ◽  
Apoptosis Proteins ◽  
Cardinal Feature

AbstractSARS-CoV-2 infection has a risk to develop into life-threatening COVID-19 disease. Whereas age, hypertension, and chronic inflammatory conditions are risk factors, underlying host factors and markers for disease severity, e.g. requiring intensive care unit (ICU) treatment, remain poorly defined. To this end, we longitudinally profiled blood inflammation markers, antibodies, and 101 plasma proteins of hospitalized COVID-19 patients who did or did not require ICU admission. While essentially all patients displayed SARS-CoV-2-specific antibodies and virus-neutralization capacity within 12–15 days, a rapid, mostly transient upregulation of selective inflammatory markers including IL-6, CXCL10, CXCL11, IFNγ, IL-10, and monocyte-attracting CCL2, CCL7 and CCL8, was particularly evident in ICU patients. In addition, there was consistent and sustained upregulation of apoptosis-associated proteins CASP8, TNFSF14, HGF, and TGFB1, with HGF discriminating between ICU and non-ICU cohorts. Thus, COVID-19 is associated with a selective inflammatory milieu within which the apoptotic pathway is a cardinal feature with potential to aid risk-based patient stratification.

scholarly journals Identification of Genes Encoding Exported Mycobacterium tuberculosis Proteins Using a Tn552′phoA In Vitro Transposition System

2000 ◽  
Vol 182 (10) ◽  
pp. 2732-2740 ◽  
Author(s):  
Miriam Braunstein ◽  
Thomas J. Griffin ◽  
Jordan I. Kriakov ◽  
Sarah T. Friedman ◽  
Nigel D. F. Grindley ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Protective Immunity ◽  
Cell Envelope ◽  
Open Reading Frames ◽  
Signal Peptides ◽  
Genomic Libraries ◽  
Target Dna ◽  
Genes Encoding ◽  
Associated Proteins

ABSTRACT Secreted and cell envelope-associated proteins are important to both Mycobacterium tuberculosis pathogenesis and the generation of protective immunity to M. tuberculosis. We used an in vitro Tn552′phoA transposition system to identify exported proteins of M. tuberculosis. The system is simple and efficient, and the transposon inserts randomly into target DNA. M. tuberculosis genomic libraries were targeted with Tn552′phoA transposons, and these libraries were screened in M. smegmatis for active PhoA translational fusions. Thirty-two different M. tuberculosis open reading frames were identified; eight contain standard signal peptides, six contain lipoprotein signal peptides, and seventeen contain one or more transmembrane domains. Four of these proteins had not yet been assigned as exported proteins in the M. tuberculosisdatabases. This collection of exported proteins includes factors that are known to participate in the immune response of M. tuberculosis and proteins with homologies, suggesting a role in pathogenesis. Nine of the proteins appear to be unique to mycobacteria and represent promising candidates for factors that participate in protective immunity and virulence. This technology of creating comprehensive fusion libraries should be applicable to other organisms.

scholarly journals Comparative Proteomic Profiling of Ehrlichia ruminantium Pathogenic Strain and Its High-Passaged Attenuated Strain Reveals Virulence and Attenuation-Associated Proteins

PLoS ONE ◽  
2015 ◽  
Vol 10 (12) ◽  
pp. e0145328 ◽  
Author(s):  
Isabel Marcelino ◽  
Miguel Ventosa ◽  
Elisabete Pires ◽  
Markus Müller ◽  
Frédérique Lisacek ◽  
...  
Keyword(s):  
Pathogenic Strain ◽  
Proteomic Profiling ◽  
Ehrlichia Ruminantium ◽  
Associated Proteins

scholarly journals Fluorescence Imaging-Based Discovery of Membrane Domain-Associated Proteins in Mycobacterium smegmatis

2021 ◽  
Author(s):  
Corelle A. Z. Rokicki ◽  
James R. Brenner ◽  
Alexander H. Dills ◽  
Julius J. Judd ◽  
Jemila C. Kester ◽  
...  
Keyword(s):  
Mycobacterium Smegmatis ◽  
Fluorescent Protein ◽  
Cell Envelope ◽  
Protein Localization ◽  
Subcellular Fractionation ◽  
Polar Regions ◽  
Intracellular Membrane ◽  
Membrane Domain ◽  
Associated Proteins ◽  
Proteomic Analyses

Mycobacteria spatially organize their plasma membrane, and many enzymes involved in envelope biosynthesis associate with a membrane compartment termed the intracellular membrane domain (IMD). The IMD is concentrated in the polar regions of growing cells and becomes less polarized under non-growing conditions. Because mycobacteria elongate from the poles, the observed polar localization of the IMD during growth likely supports the localized biosynthesis of envelope components. While we have identified more than 300 IMD-associated proteins by proteomic analyses, only a handful of these have been verified by independent experimental methods. Furthermore, some IMD-associated proteins may have escaped proteomic identification and remain to be identified. Here, we visually screened an arrayed library of 523 Mycobacterium smegmatis strains, each producing a Dendra2-FLAG-tagged recombinant protein. We identified 29 fusion proteins that showed polar fluorescence patterns characteristic of IMD proteins. Twenty of these had previously been suggested to localize to the IMD based on proteomic data. Of the nine remaining IMD candidate proteins, three were confirmed by biochemical methods to be associated with the IMD. Taken together, this new co-localization strategy is effective in verifying the IMD association of proteins found by proteomic analyses, while facilitating the discovery of additional IMD-associated proteins. Importance The intracellular membrane domain (IMD) is a membrane subcompartment found in Mycobacterium smegmatis cells. Proteomic analysis of purified IMD identified more than 300 proteins, including enzymes involved in cell envelope biosynthesis. However, proteomics on its own is unlikely to detect every IMD-associated protein because of technical and biological limitations. Here, we describe fluorescent protein co-localization as an alternative, independent approach. Using a combination of fluorescence microscopy, proteomics, and subcellular fractionation, we identified three new proteins associated with the IMD. Such a robust method to rigorously define IMD proteins will benefit future investigations to decipher the synthesis, maintenance and functions of this membrane domain, and help delineate a more general mechanisms of subcellular protein localization in mycobacteria.

scholarly journals Definition of novel cell envelope associated proteins in Triton X-114 extracts of Mycobacterium tuberculosis H37Rv

2010 ◽  
Vol 10 (1) ◽  
pp. 132 ◽  
Author(s):  
Hiwa Målen ◽  
Sharad Pathak ◽  
Tina Søfteland ◽  
Gustavo A de Souza ◽  
Harald G Wiker
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Cell Envelope ◽  
Tuberculosis H37rv ◽  
Mycobacterium Tuberculosis H37rv ◽  
Associated Proteins ◽  
Definition Of ◽  
Triton X

scholarly journals The cell-envelope proteome of Bifidobacterium longum in an in vitro bile environment

Microbiology ◽  
2009 ◽  
Vol 155 (3) ◽  
pp. 957-967 ◽  
Author(s):  
Lorena Ruiz ◽  
Yohann Couté ◽  
Borja Sánchez ◽  
Clara G. de los Reyes-Gavilán ◽  
Jean-Charles Sanchez ◽  
...  
Keyword(s):  
Membrane Fraction ◽  
Ribosomal Proteins ◽  
Soluble Fraction ◽  
Cell Envelope ◽  
Bifidobacterium Longum ◽  
Cell Wall Proteins ◽  
Stable Isotope Labelling ◽  
Cytoplasmic Proteins ◽  
Associated Proteins

Host–bacteria interactions are often mediated via surface-associated proteins. The identification of these proteins is an important goal of bacterial proteomics. To address how bile can influence the cell-envelope proteome of Bifidobacterium longum biotype longum NCIMB 8809, we analysed its membrane protein fraction using stable isotope labelling of amino acids in cell culture (SILAC). We were able to identify 141 proteins in the membrane fraction, including a large percentage of the theoretical transporters of this species. Moreover, the envelope-associated soluble fraction was analysed using different subfractionation techniques and differential in-gel fluorescence electrophoresis (DIGE). This approach identified 128 different proteins. Some of them were well-known cell wall proteins, but others were highly conserved cytoplasmic proteins probably displaying a ‘moonlighting’ function. We were able to identify 11 proteins in the membrane fraction and 6 proteins in the envelope-associated soluble fraction whose concentration varied in the presence of bile. Bile promoted changes in the levels of proteins with important biological functions, such as some ribosomal proteins and enolase. Also, oligopeptide-binding proteins were accumulated on the cell surface, which was reflected in a different tripeptide transport rate in the cells grown with bile. The data reported here will provide the first cell-envelope proteome map for B. longum, and may contribute to understanding the bile tolerance of these bacteria.

scholarly journals Proteomic profiling of Myc-associated proteins 

Cell Cycle ◽  
2010 ◽  
Vol 9 (24) ◽  
pp. 4908-4921 ◽  
Author(s):  
Pooja Agrawal ◽  
Kebing Yu ◽  
Arthur R. Salomon ◽  
John M. Sedivy
Keyword(s):  
Proteomic Profiling ◽  
Associated Proteins
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