Fe(iii)-complex mediated bacterial cell surface immobilization of eGFP and enzymes

2021 ◽  
Author(s):  
Lilin Feng ◽  
Liang Gao ◽  
Daniel F. Sauer ◽  
Yu Ji ◽  
Haiyang Cui ◽  
...  
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Surface Immobilization ◽  
E Coli ◽  
Bacterial Cell Surface ◽  
Reversible Method

A facile and reversible method to immobilize His6-tagged proteins on the E. coli cell surface through the formation of an Fe(iii)-complex.

scholarly journals Heterogeneous Surface Expression of EspA Translocon Filaments by Escherichia coli O157:H7 Is Controlled at the Posttranscriptional Level

2003 ◽  
Vol 71 (10) ◽  
pp. 5900-5909 ◽  
Author(s):  
Andrew J. Roe ◽  
Helen Yull ◽  
Stuart W. Naylor ◽  
Martin J. Woodward ◽  
David G. E. Smith ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Bacterial Population ◽  
Enteric Bacteria ◽  
Effector Proteins ◽  
Host Cells ◽  
Phase Variable ◽  
E Coli ◽  
Bacterial Cell Surface

ABSTRACT Type III secretion systems of enteric bacteria enable translocation of effector proteins into host cells. Secreted proteins of verotoxigenic Escherichia coli O157 strains include components of a translocation apparatus, EspA, -B, and -D, as well as “effectors” such as the translocated intimin receptor (Tir) and the mitochondrion-associated protein (Map). This research has investigated the regulation of LEE4 translocon proteins, in particular EspA. EspA filaments could not be detected on the bacterial cell surface when E. coli O157:H7 was cultured in M9 minimal medium but were expressed from only a proportion of the bacterial population when cultured in minimal essential medium modified with 25 mM HEPES. The highest proportions of EspA-filamented bacteria were detected in late exponential phase, after which filaments were lost rapidly from the bacterial cell surface. Our previous research had shown that human and bovine E. coli O157:H7 strains exhibit marked differences in EspD secretion levels. Here it is demonstrated that the proportion of the bacterial population expressing EspA filaments was associated with the level of EspD secretion. The ability of individual bacteria to express EspA filaments was not controlled at the level of LEE1-4 operon transcription, as demonstrated by using both β-galactosidase and green fluorescent protein (GFP) promoter fusions. All bacteria, whether expressing EspA filaments or not, showed equivalent levels of GFP expression when LEE1-4 translational fusions were used. Despite this, the LEE4-espADB mRNA was more abundant from populations with a high proportion of nonsecreting bacteria (low secretors) than from populations with a high proportion of secreting and therefore filamented bacteria (high secretors). This research demonstrates that while specific environmental conditions are required to induce LEE1-4 expression, a further checkpoint exists before EspA filaments are produced on the bacterial surface and secretion of effector proteins occurs. This checkpoint in E. coli O157:H7 translocon expression is controlled by a posttranscriptional mechanism acting on LEE4-espADB mRNA. The heterogeneity in EspA filamentation could arise from phase-variable expression of regulators that control this posttranscriptional mechanism.

scholarly journals Four Different Genes Responsible for Nonimmune Immunoglobulin-Binding Activities within a Single Strain ofEscherichia coli

2000 ◽  
Vol 68 (4) ◽  
pp. 2205-2214 ◽  
Author(s):  
Carol H. Sandt ◽  
Charles W. Hill
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Outer Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Single Strain ◽  
E Coli ◽  
Bacterial Cell Surface ◽  
Igg Binding ◽  
K 12

ABSTRACT Certain Escherichia coli strains bind the Fc fragment of immunoglobulin G (IgG) at the bacterial cell surface. Previous work established that this nonimmune Ig binding depends on several large proteins with apparent molecular masses that can exceed 200 kDa. ForE. coli strain ECOR-9, four distinct genes (designatedeibA, eibC, eibD, andeibE) are responsible for Ig binding. Two eibgenes are linked to eaa genes, which are homologous to genes for the autotransporter family of secreted proteins. With reference to the E. coli K-12 chromosome, theeibA-eaaA cluster is adjacent to trpA (min 28.3) while the eibC-eaaC cluster is adjacent toaspS (min 42.0). Sequence adjacent to theeibA-eaaA cluster converges with that of strain K-12 precisely as observed for the Atlas family of prophages, suggesting that eibA is part of one of these. All four eibgenes, when cloned into plasmid vectors, impart IgG binding to E. coli K-12 strains, and three impart IgA binding also. The IgG binding occurs at the bacterial cell surface, and its expression increases survival in serum by up to 3 orders of magnitude. Theeib sequences predict a C-terminal peptide motif that is characteristic of outer membrane proteins, and the protein sequences show significant similarity near the C terminus to both the YadA virulence factor of Yersinia species and the universal surface protein A II of Moraxella catarrhalis. The sizes predicted for Eib proteins from DNA sequence are much smaller than their apparent sizes on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, possibly reflecting stable oligomerization.

scholarly journals Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy

The Analyst ◽  
2014 ◽  
Vol 139 (12) ◽  
pp. 3174-3178 ◽  
Author(s):  
Ian L. Gunsolus ◽  
Dehong Hu ◽  
Cosmin Mihai ◽  
Samuel E. Lohse ◽  
Chang-soo Lee ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Super Resolution ◽  
Bacterial Cell Surface

The Bacterial Cell Surface in Relation to the Environment

1984 ◽  
pp. 194-219
Author(s):  
Stephen M. Hammond ◽  
Peter A. Lambert ◽  
Andrew N. Rycroft
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Bacterial Cell Surface

scholarly journals Cover Feature: Bacterial Cell‐Surface Display of Semisynthetic Cyclic Peptides (ChemBioChem 1/2019)

ChemBioChem ◽  
2018 ◽  
Vol 20 (1) ◽  
pp. 4-4
Author(s):  
Shubhendu Palei ◽  
Kira S. Becher ◽  
Christian Nienberg ◽  
Joachim Jose ◽  
Henning D. Mootz
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Cyclic Peptides ◽  
Surface Display ◽  
Cell Surface Display ◽  
Bacterial Cell Surface

scholarly journals Spin-Dependent Electron Transport through Bacterial Cell Surface Multiheme Electron Conduits

2019 ◽  
Author(s):  
Suryakant Mishra ◽  
Sahand Pirbadian ◽  
Amit Kumar Mondal ◽  
Moh El-Naggar ◽  
Ron Naaman
Keyword(s):  
Electron Transport ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Shewanella Oneidensis ◽  
Extracellular Electron Transfer ◽  
Long Distance ◽  
Force Microscopy ◽  
Bacterial Cell Surface ◽  
Redox Active ◽  
Gain Energy

Multiheme cytochromes, located on the bacterial cell surface, function as long-distance (> 10 nm) electron conduits linking intracellular reactions to external surfaces. This extracellular electron transfer process, which allows microorganisms to gain energy by respiring solid redox-active minerals, also facilitates the wiring of cells to electrodes. While recent studies suggested that a chiral induced spin selectivity effect is linked to efficient electron transmission through biomolecules, this phenomenon has not been investigated in the extracellular electron conduits. Using magnetic conductive probe atomic force microscopy, Hall voltage measurements, and spin-dependent electrochemistry of the decaheme cytochromes MtrF and OmcA from the metal-reducing bacterium <i>Shewanella oneidensis</i> MR-1, we show that electron transport through these extracellular conduits is spin-selective. Our study has implications for understanding how spin-dependent interactions and magnetic fields may control electron transport across biotic-abiotic interfaces in both natural and biotechnological systems.

scholarly journals Editorial: The many wonders of the bacterial cell surface

2015 ◽  
Vol 40 (2) ◽  
pp. 161-163 ◽  
Author(s):  
Alain Filloux ◽  
Chris Whitfield
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Bacterial Cell Surface ◽  
The Many

scholarly journals Phosphoethanolamine cellulose enhances curli-mediated adhesion of uropathogenicEscherichia colito bladder epithelial cells

2018 ◽  
Vol 115 (40) ◽  
pp. 10106-10111 ◽  
Author(s):  
Emily C. Hollenbeck ◽  
Alexandra Antonoplis ◽  
Chew Chai ◽  
Wiriya Thongsomboon ◽  
Gerald G. Fuller ◽  
...  
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Cell Monolayer ◽  
Surface Structures ◽  
Bacterial Cell Surface ◽  
Cell Surface Structures ◽  
Bladder Cell ◽  
Type 1 Pili ◽  
Tract Infections

UropathogenicEscherichia coli(UPEC) are the major causative agents of urinary tract infections, employing numerous molecular strategies to contribute to adhesion, colonization, and persistence in the bladder niche. Identifying strategies to prevent adhesion and colonization is a promising approach to inhibit bacterial pathogenesis and to help preserve the efficacy of available antibiotics. This approach requires an improved understanding of the molecular determinants of adhesion to the bladder urothelium. We designed experiments using a custom-built live cell monolayer rheometer (LCMR) to quantitatively measure individual and combined contributions of bacterial cell surface structures [type 1 pili, curli, and phosphoethanolamine (pEtN) cellulose] to bladder cell adhesion. Using the UPEC strain UTI89, isogenic mutants, and controlled conditions for the differential production of cell surface structures, we discovered that curli can promote stronger adhesive interactions with bladder cells than type 1 pili. Moreover, the coproduction of curli and pEtN cellulose enhanced adhesion. The LCMR enables the evaluation of adhesion under high-shear conditions to reveal this role for pEtN cellulose which escaped detection using conventional tissue culture adhesion assays. Together with complementary biochemical experiments, the results support a model wherein cellulose serves a mortar-like function to promote curli association with and around the bacterial cell surface, resulting in increased bacterial adhesion strength at the bladder cell surface.

Sign in / Sign up

Export Citation Format

Share Document