scholarly journals A general approach to explore prokaryotic protein glycosylation reveals the unique surface layer modulation of an anammox bacterium

2021 ◽  
Author(s):  
Martin Pabst ◽  
Denis S. Grouzdev ◽  
Christopher E. Lawson ◽  
Hugo B. C. Kleikamp ◽  
Carol de Ram ◽  
...  
Keyword(s):  
Surface Layer ◽  
Protein Glycosylation ◽  
Anammox Bacterium ◽  
Prokaryotic Protein ◽  
Unique Surface

scholarly journals A general approach to explore prokaryotic protein glycosylation reveals the unique surface layer modulation of an anammox bacterium

2020 ◽  
Author(s):  
Martin Pabst ◽  
Denis Grouzdev ◽  
Christopher E. Lawson ◽  
Hugo B.C. Kleikamp ◽  
Carol de Ram ◽  
...  
Keyword(s):  
Surface Layer ◽  
Large Scale ◽  
Enrichment Culture ◽  
Biological Significance ◽  
Protein Glycosylation ◽  
Chemical Diversity ◽  
Proteomics Data ◽  
Cellular Protection ◽  
Evolutionary Forces ◽  
Prokaryotic Protein

The enormous chemical diversity and strain variability of prokaryotic protein glycosylation makes a large-scale exploration exceptionally challenging. Therefore, despite the universal relevance of protein glycosylation across all domains of life, the understanding of their biological significance and the evolutionary forces shaping oligosaccharide structures remains highly limited.Here, we report on a newly established mass binning glycoproteomics approach that establishes the chemical identity of the carbohydrate components and performs untargeted exploration of prokaryotic oligosaccharides from large-scale proteomics data directly. We demonstrate our approach by exploring an enrichment culture of the globally relevant anaerobic ammonium-oxidizing bacterium Ca. Kuenenia stuttgartiensis. By doing so we resolved a remarkable array of oligosaccharides, produced by two entirely unrelated glycosylation machineries targeting the same surface-layer protein (SLP) simultaneously. More intriguingly, the investigated strain also accomplished modulation of highly specialized sugars, supposedly in response to its energy metabolism—the anaerobic oxidation of ammonium —which depends on the acquisition of substrates of opposite charge. Ultimately, we provide a systematic approach for the compositional exploration of prokaryotic protein glycosylation, and reveal for the first time a remarkable balance between maximising cellular protection through a complex array of oligosaccharides and adhering to the requirements of the ‘metabolic lifestyle’.

scholarly journals Identification of the Major Expressed S-Layer and Cell Surface-Layer-Related Proteins in the Model Methanogenic Archaea:Methanosarcina barkeriFusaro andMethanosarcina acetivoransC2A

Archaea ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Lars Rohlin ◽  
Deborah R. Leon ◽  
Unmi Kim ◽  
Joseph A. Loo ◽  
Rachel R. Ogorzalek Loo ◽  
...  
Keyword(s):  
Surface Layer ◽  
Cell Surface ◽  
Structural Integrity ◽  
Lipid Membrane ◽  
Expression Patterns ◽  
Methanosarcina Barkeri ◽  
Protein Glycosylation ◽  
Methanosarcina Acetivorans ◽  
Genome Annotations ◽  
Cell Envelopes

Many archaeal cell envelopes contain a protein coat or sheath composed of one or more surface exposed proteins. These surface layer (S-layer) proteins contribute structural integrity and protect the lipid membrane from environmental challenges. To explore the species diversity of these layers in the Methanosarcinaceae, the major S-layer protein inMethanosarcina barkeristrain Fusaro was identified using proteomics. The Mbar_A1758 gene product was present in multiple forms with apparent sizes of 130, 120, and 100 kDa, consistent with post-translational modifications including signal peptide excision and protein glycosylation. A protein with features related to the surface layer proteins found inMethanosarcina acetivoransC2A andMethanosarcina mazeiGoel was identified in theM. barkerigenome. These data reveal a distinct conserved protein signature with features and implied cell surface architecture in the Methanosarcinaceae that is absent in other archaea. Paralogous gene expression patterns in twoMethanosarcinaspecies revealed abundant expression of a single S-layer paralog in each strain. Respective promoter elements were identified and shown to be conserved in mRNA coding and upstream untranslated regions. PriorM. acetivoransgenome annotations assigned S-layer or surface layer associated roles of eighty genes: however, of 68 examined none was significantly expressed relative to the experimentally determined S-layer gene.

scholarly journals Comparative Analysis of Surface Layer Glycoproteins and Genes Involved in Protein Glycosylation in the Genus Haloferax

Genes ◽  
2018 ◽  
Vol 9 (3) ◽  
pp. 172 ◽  
Author(s):  
Yarden Shalev ◽  
Shannon Soucy ◽  
R. Papke ◽  
J. Gogarten ◽  
Jerry Eichler ◽  
...  
Keyword(s):  
Surface Layer ◽  
Comparative Analysis ◽  
Protein Glycosylation

The oxidation of Inconel alloy MA754 at low oxidation potential

1983 ◽  
Vol 41 ◽  
pp. 218-219
Author(s):  
D. N. Braski ◽  
P. D. Goodell ◽  
J. V. Cathcart ◽  
R. H. Kane
Keyword(s):  
Surface Layer ◽  
Oxidation Potential ◽  
Metal Interface ◽  
Elevated Temperature Strength ◽  
Surface Conditions ◽  
Inconel Alloy ◽  
Inco Alloy ◽  
Resistance To Oxidation ◽  
Oxide Particles ◽  
Cold Worked

It has been known for some time that the addition of small oxide particles to an 80 Ni—20 Cr alloy not only increases its elevated-temperature strength, but also markedly improves its resistance to oxidation. The mechanism by which the oxide dispersoid enhances the oxidation resistance is being studied collaboratively by ORNL and INCO Alloy Products Company.Initial experiments were performed using INCONEL alloy MA754, which is nominally: 78 Ni, 20 Cr, 0.05 C, 0.3 Al, 0.5 Ti, 1.0 Fe, and 0.6 Y2O3 (wt %).Small disks (3 mm diam × 0.38 mm thick) were cut from MA754 plate stock and prepared with two different surface conditions. The first was prepared by mechanically polishing one side of a disk through 0.5 μm diamond on a syntron polisher while the second used an additional sulfuric acid-methanol electropolishing treatment to remove the cold-worked surface layer. Disks having both surface treatments were oxidized in a radiantly heated furnace for 30 s at 1000°C. Three different environments were investigated: hydrogen with nominal dew points of 0°C, —25°C, and —55°C. The oxide particles and films were examined in TEM by using extraction replicas (carbon) and by backpolishing to the oxide/metal interface. The particles were analyzed by EDS and SAD.

Structural analysis of the surface-layer protein of spirillum serpens by high-resolution electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 738-739
Author(s):  
W. H. Wu ◽  
R. M. Glaeser
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Structural Analysis ◽  
High Resolution ◽  
Low Dose ◽  
High Resolution Electron Microscopy ◽  
Optical Diffraction ◽  
Surface Layer Protein ◽  
Morphological Unit ◽  
Resolution Electron

Spirillum serpens possesses a surface layer protein which exhibits a regular hexagonal packing of the morphological subunits. A morphological model of the structure of the protein has been proposed at a resolution of about 25 Å, in which the morphological unit might be described as having the appearance of a flared-out, hollow cylinder with six ÅspokesÅ at the flared end. In order to understand the detailed association of the macromolecules, it is necessary to do a high resolution structural analysis. Large, single layered arrays of the surface layer protein have been obtained for this purpose by means of extensive heating in high CaCl2, a procedure derived from that of Buckmire and Murray. Low dose, low temperature electron microscopy has been applied to the large arrays.As a first step, the samples were negatively stained with neutralized phosphotungstic acid, and the specimens were imaged at 40,000 magnification by use of a high resolution cold stage on a JE0L 100B. Low dose images were recorded with exposures of 7-9 electrons/Å2. The micrographs obtained (Fig. 1) were examined by use of optical diffraction (Fig. 2) to tell what areas were especially well ordered.

Electron-Mirror Microscopic Aspects of Ferroelectric Domains of BaTiO3 And Ca2Sr (C2H5CO2)6

1970 ◽  
Vol 28 ◽  
pp. 478-479
Author(s):  
Teruo Someya ◽  
Jinzo Kobayashi
Keyword(s):  
Surface Layer ◽  
Electric Fields ◽  
Quantitative Study ◽  
Recent Progress ◽  
Ferroelectric Domain ◽  
Resolving Power ◽  
Surface Pattern ◽  
Crystal Surfaces ◽  
One Dimensional ◽  
Ferroelectric Domains

Recent progress in the electron-mirror microscopy (EMM), e.g., an improvement of its resolving power together with an increase of the magnification makes it useful for investigating the ferroelectric domain physics. English has recently observed the domain texture in the surface layer of BaTiO3. The present authors ) have developed a theory by which one can evaluate small one-dimensional electric fields and/or topographic step heights in the crystal surfaces from their EMM pictures. This theory was applied to a quantitative study of the surface pattern of BaTiO3).

Microstructure of Electro-Eroded Cemented Carbide Surfaces

1968 ◽  
Vol 26 ◽  
pp. 284-285
Author(s):  
V. N. Filimonenko ◽  
M. H. Richman ◽  
J. Gurland
Keyword(s):  
Surface Layer ◽  
Cobalt Content ◽  
Cemented Carbides ◽  
Face Centered Cubic ◽  
X Ray Diffraction ◽  
Metallographic Examination ◽  
Standard Procedures ◽  
Face Centered ◽  
Diamond Paste ◽  
Plastic Carbon

The high temperatures and pressures that are found in a spark gap during electrical discharging lead to a sharp phase transition and structural transformation in the surface layer of cemented carbides containing WC and cobalt. By means of X-ray diffraction both W2C and a high-temperature monocarbide of tungsten (face-centered cubic) were detected after electro-erosion. The W2C forms as a result of the peritectic reaction, WC → W2C+C. The existence and amount of the phases depend on both the energy of the electro-spark discharge and the cobalt content. In the case of a low-energy discharge (i.e. C=0.01μF, V = 300v), WC(f.c.c.) is generally formed in the surface layer. However, at high energies, (e.g. C=30μF, V = 300v), W2C is formed at the surface in preference to the monocarbide. The phase transformations in the surface layer are retarded by the presence of larger percentages of cobalt.Metallographic examination of the electro-eroded surfaces of cemented carbides was carried out on samples with 5-30% cobalt content. The specimens were first metallographically polished using diamond paste and standard procedures and then subjected to various electrical discharges on a Servomet spark machining device. The samples were then repolished and etched in a 3% NH4OH electrolyte at -0.5 amp/cm2. Two stage plastic-carbon replicas were then made and shadowed with chromium at 27°.

Effect of brefeldin A (BFA) on the unique surface coat of pulmonary intravascular macrophages (PIMs) of sheep: Ultrastructural and cytochemical study

1993 ◽  
Vol 51 ◽  
pp. 18-19
Author(s):  
Baljit Singh
Keyword(s):  
Brefeldin A ◽  
Ultrastructural Feature ◽  
Surface Coat ◽  
Anterograde Transport ◽  
Cytochemical Study ◽  
Active Involvement ◽  
Electron Lucent ◽  
Pulmonary Intravascular Macrophages ◽  
Unique Surface

The PIM of sheep, calf, goat and horse has a characteristic ultrastructural feature in the form of a unique, heparin sensitive, globular surface coat present around the plasma membrane with an intervening electron lucent space of 32-40 nm. We previously showed the active involvement of this surface coat in the phagocytosis of tracer material like monastral blue and cationized ferritin. The surface coat is capable of reconstitution in vivo following disruption with heparin. The present study was aimed to investigate whether PIM is the source of surface coat or not. In the recent years the BFA has been extensively used to understand the secretory pathways in the cells because of its ability to cause a rapid and reversible block to the anterograde transport of proteins from the endoplasmic reticulum to the Golgi.Sheep (n=6) were weighed, their plasma volume was calculated indirectly and based on which a sufficient single intravenous dose of BFA was given so as to reach a concentration of 4-5 microgram/ml of plasma.

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