scholarly journals Functional characterization of the FimH adhesin from Salmonella enterica serovar Enteritidis

Microbiology ◽  
2006 ◽  
Vol 152 (5) ◽  
pp. 1337-1346 ◽  
Author(s):  
Dagmara Kisiela ◽  
Anna Laskowska ◽  
Anna Sapeta ◽  
Maciej Kuczkowski ◽  
Alina Wieliczko ◽  
...  
Keyword(s):  
Horseradish Peroxidase ◽  
Salmonella Enterica ◽  
Functional Characterization ◽  
Infectious Agent ◽  
Binding Specificity ◽  
Human Colon ◽  
Binding Properties ◽  
Intact Cells ◽  
Serovar Typhimurium ◽  
Rnase B

Salmonella enterica serovar Enteritidis has emerged during the last 20 years as the major causative agent of food-borne gastroenteritis in humans and as the major infectious agent on poultry farms, replacing Salmonella enterica serovar Typhimurium as the dominant pathogenic serovar. Because adhesion to gut tissues and colonization of the alimentary tract, mediated in large part by the FimH adhesins located on type 1 fimbriae, is an important stage in the pathogenesis of both serovars, the binding properties of the FimH adhesins from these two enteropathogens were compared. Salmonella Enteritidis FimH protein and the Salmonella Typhimurium low-adhesive variant of this adhesin were expressed in Escherichia coli and the recombinant proteins were analysed for their ability to bind glycoproteins carrying different oligomannosidic structures and different types of eukaryotic cells. In static binding assays (ELISA and Western blotting) both FimH proteins bound equally well to all three tested glycoproteins (RNase B, horseradish peroxidase and mannan-BSA). In addition, no differences were found in the binding specificity of the FimH proteins and intact cells of Salmonella Enteritidis and Salmonella Typhimurium to human colon carcinoma or bladder cancer cells. The presence of the same amino acid residues at positions 61 (glycine) and 118 (phenylalanine) and the similar binding properties of these two adhesins suggest that the newly described FimH protein of Salmonella Enteritidis represents the low-adhesive variant found in Salmonella Typhimurium. To study the binding specificity of Salmonella Enteritidis FimH protein further, direct kinetic analysis using surface plasmon resonance was performed. With this method it was found that Salmonella Enteritidis FimH adhesin bound with the highest K d value to high-mannose type N-glycans carried by RNase B; about 100 times lower K d values were obtained in the interactions with mannan-BSA and horseradish peroxidase.

scholarly journals Characterization of FimH Adhesins Expressed by Salmonella enterica Serovar Gallinarum Biovars Gallinarum and Pullorum: Reconstitution of Mannose-Binding Properties by Single Amino Acid Substitution

2005 ◽  
Vol 73 (9) ◽  
pp. 6187-6190 ◽  
Author(s):  
Dagmara Kisiela ◽  
Anna Sapeta ◽  
Maciej Kuczkowski ◽  
Tadeusz Stefaniak ◽  
Alina Wieliczko ◽  
...  
Keyword(s):  
Colon Carcinoma ◽  
Salmonella Enterica ◽  
Human Colon ◽  
Site Directed Mutagenesis ◽  
Single Amino Acid ◽  
Binding Properties ◽  
Type 1 Fimbriae ◽  
Serovar Typhimurium ◽  
High Mannose

ABSTRACT Recombinant FimH adhesins of type 1 fimbriae from Salmonella enterica serovar Gallinarum biovars Gallinarum and Pullorum, in contrast to those of Salmonella enterica serovar Typhimurium, did not bind to high-mannose oligosaccharides or to human colon carcinoma HT-29 cells. However, mutated FimH proteins from biovar Gallinarum and biovar Pullorum, in which the isoleucine at position 78 was replaced by the threonine found in S. enterica serovar Typhimurium, bound well to glycoproteins carrying high-mannose oligosaccharides and colon carcinoma cells. The loss of sugar-binding properties by biovar Gallinarum and biovar Pullorum FimH adhesins, which are a part of the type 1 fimbriae, is most probably the result of a single T78I mutation, as was proven by site-directed mutagenesis of FimH proteins.

scholarly journals Structural and Functional Characterization of ScsC, a Periplasmic Thioredoxin-Like Protein from Salmonella enterica Serovar Typhimurium

2013 ◽  
Vol 19 (13) ◽  
pp. 1494-1506 ◽  
Author(s):  
Mark Shepherd ◽  
Begoña Heras ◽  
Maud E. S. Achard ◽  
Gordon J. King ◽  
M. Pilar Argente ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Functional Characterization ◽  
Serovar Typhimurium

scholarly journals Antimicrobial and Antivirulence Impacts of Phenolics on Salmonella Enterica Serovar Typhimurium

Antibiotics ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 668
Author(s):  
Zabdiel Alvarado-Martinez ◽  
Paulina Bravo ◽  
Nana-Frekua Kennedy ◽  
Mayur Krishna ◽  
Syed Hussain ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Infectious Agent ◽  
Resistance Pattern ◽  
Fluorescent Intensity ◽  
Antimicrobial Potential ◽  
Serovar Typhimurium ◽  
Morphological Defects ◽  
The Usa ◽  
Alternative Antimicrobials

Salmonella enterica serovar Typhimurium (ST) remains a major infectious agent in the USA, with an increasing antibiotic resistance pattern, which requires the development of novel antimicrobials capable of controlling ST. Polyphenolic compounds found in plant extracts are strong candidates as alternative antimicrobials, particularly phenolic acids such as gallic acid (GA), protocatechuic acid (PA) and vanillic acid (VA). This study evaluates the effectiveness of these compounds in inhibiting ST growth while determining changes to the outer membrane through fluorescent dye uptake and scanning electron microscopy (SEM), in addition to measuring alterations to virulence genes with qRT-PCR. Results showed antimicrobial potential for all compounds, significantly inhibiting the detectable growth of ST. Fluorescent spectrophotometry and microscopy detected an increase in relative fluorescent intensity (RFI) and red-colored bacteria over time, suggesting membrane permeabilization. SEM revealed severe morphological defects at the polar ends of bacteria treated with GA and PA, while VA-treated bacteria were found to be mid-division. Relative gene expression showed significant downregulation in master regulator hilA and invH after GA and PA treatments, while fliC was upregulated in VA. Results suggest that GA, PA and VA have antimicrobial potential that warrants further research into their mechanism of action and the interactions that lead to ST death.

scholarly journals Potential Involvement of Salmonella Infection in Autoimmunity

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 96 ◽  
Author(s):  
Zhanna Ktsoyan ◽  
Lyudmila Budaghyan ◽  
Marina Agababova ◽  
Armine Mnatsakanyan ◽  
Karine Arakelova ◽  
...  
Keyword(s):  
Autoimmune Disease ◽  
Post Infection ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Profile Analysis ◽  
Infectious Agent ◽  
Disease Stage ◽  
Salmonella Infection ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Serovar Typhimurium

In this work, we investigated the potential effects of nontyphoidal Salmonella infection on autoantibody (AA) formation. The titer and profiles of autoantibodies in the sera of patients with acute salmonellosis due to Salmonella enterica serovar Typhimurium (S. Typhimurium) or Salmonella enterica serovar Enteritidis (S. Enteritidis) infection, as well as in convalescent patients, were determined with indirect immunofluorescence. A significant increase of autoantibodies in acute diseases caused by both serotypes of Salmonella and during post infection by S. Enteritidis was detected. Antibody profile analysis by multivariate statistics revealed that this increase was non-specific and was not dependent on the infectious agent or disease stage. The results obtained suggest that nontyphoidal Salmonella infection contributes to the generation of autoantibodies and may play a role in autoimmune disease.

scholarly journals Identification and Functional Characterization of Chicken Toll-Like Receptor 5 Reveals a Fundamental Role in the Biology of Infection with Salmonella enterica Serovar Typhimurium

2005 ◽  
Vol 73 (4) ◽  
pp. 2344-2350 ◽  
Author(s):  
Muhammad Iqbal ◽  
Victoria J. Philbin ◽  
G. S. K. Withanage ◽  
Paul Wigley ◽  
Richard K. Beal ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Functional Characterization ◽  
Systemic Infection ◽  
Structural Similarity ◽  
Conserved Sequence ◽  
Receptor Repertoire ◽  
Serovar Typhimurium ◽  
Toll Like Receptor 5

ABSTRACT Toll-like receptors (TLRs) are a major component of the pattern recognition receptor repertoire that detect invading microorganisms and direct the vertebrate immune system to eliminate infection. In chickens, the differential biology of Salmonella serovars (systemic versus gut-restricted localization) correlates with the presence or absence of flagella, a known TLR5 agonist. Chicken TLR5 (chTLR5) exhibits conserved sequence and structural similarity with mammalian TLR5 and is expressed in tissues and cell populations of immunological and stromal origin. Exposure of chTLR5+ cells to flagellin induced elevated levels of chicken interleukin-1β (chIL-1β) but little upregulation of chIL-6 mRNA. Consistent with the flagellin-TLR5 hypothesis, an aflagellar Salmonella enterica serovar Typhimurium fliM mutant exhibited an enhanced ability to establish systemic infection. During the early stages of infection, the fliM mutant induced less IL-1β mRNA and polymorphonuclear cell infiltration of the gut. Collectively, the data represent the identification and functional characterization of a nonmammalian TLR5 and indicate a role in restricting the entry of flagellated Salmonella into systemic sites of the chicken.

scholarly journals Structural and functional characterization of Salmonella enterica serovar Typhimurium YcbL: An unusual Type II glyoxalase

2010 ◽  
Vol 19 (10) ◽  
pp. 1897-1905 ◽  
Author(s):  
Anna L. Stamp ◽  
Paul Owen ◽  
Kamel El Omari ◽  
Charles E. Nichols ◽  
Michael Lockyer ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Functional Characterization ◽  
Type Ii ◽  
Serovar Typhimurium ◽  
Unusual Type

scholarly journals The C-Terminal Domain of Salmonella enterica Serovar Typhimurium OmpA Is an Immunodominant Antigen in Mice but Appears To Be Only Partially Exposed on the Bacterial Cell Surface

2003 ◽  
Vol 71 (7) ◽  
pp. 3937-3946 ◽  
Author(s):  
Shiva P. Singh ◽  
Yvonne U. Williams ◽  
Stephanie Miller ◽  
Hiroshi Nikaido
Keyword(s):  
Cell Surface ◽  
Outer Membrane ◽  
Bacterial Cell ◽  
Salmonella Enterica ◽  
Competitive Inhibition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Intact Cells ◽  
Bacterial Cell Surface ◽  
Terminal Domain ◽  
Serovar Typhimurium

ABSTRACT We examined the way the major outer membrane protein OmpA of Salmonella enterica serovar Typhimurium is recognized by the mouse immune system, by raising a panel of 12 monoclonal antibodies (MAbs) against this protein. Interaction between OmpA and these MAbs is competitively inhibited with several-hundredfold dilutions of mouse polyclonal sera obtained by immunization with live or heat-killed whole cells, suggesting that OmpA is one of the immunodominant antigens of serovar Typhimurium. All of the MAbs were specific for an identical epitope(s) located on the C-terminal domain of OmpA, as indicated by the use of OmpA fragments generated by protease or cyanogen bromide treatment and by competitive inhibition enzyme-linked immunosorbent assay. This epitope was highly conserved within (but not outside) the family Enterobacteriaceae. The strong immunogenicity of this epitope was surprising because the C-terminal domain of OmpA, usually thought to be located in the periplasm, is not expected to be exposed on the bacterial cell surface. A MAb, however, reacted in a cytofluorometry assay more strongly with outer-membrane-permeabilized cells than with untreated cells, a result supporting the predominantly periplasmic localization of the epitope. Significant, though low-level, reactivity of intact cells nevertheless suggests that in some cells the C-terminal domain of OmpA is exposed on the surface, a result consistent with the proposal that OmpA can fold into one of the two alternate conformations.

scholarly journals Whole genome analysis and characterization of Salmonella enterica subsp. enterica serovar Typhimurium strain UPM 260 for mediated genetic manipulation

2020 ◽  
Author(s):  
Najwa Syahirah Roslan ◽  
Nurulfiza Mat Isa ◽  
Abdul Rahman Omar ◽  
Mohd. Hair Bejo ◽  
Aini Ideris
Keyword(s):  
Salmonella Enterica ◽  
Functional Characterization ◽  
Poultry Meat ◽  
Whole Genome Sequence ◽  
Health Concern ◽  
Type I ◽  
Whole Genome ◽  
Typhimurium Strain ◽  
Serovar Typhimurium ◽  
The Impact

Abstract Background Salmonella enterica serovar Typhimurium persists as one of the most frequent food-borne zoonoses, causing a major public health concern worldwide. Furthermore, Salmonella infection has a large economic impact. Globally, the main sources of infection for humans include the consumption of contaminated poultry meat and eggs. In animals however, Salmonella transmission usually occurs horizontally from infected birds and contaminated environments. Hence, to delve further on how the impact of this disease can be lessened, an epidemiological study needs to be performed. It is vital to determine the genomic sequences of microorganisms to understand their biology and functional characterization. Thus, we determined the whole-genome sequence and virulence profile of S. enterica serovar Typhimurium strain UPM 260 isolated from Perak, Malaysia. Whole genome sequencing (WGS) using paired-end sequencing generated 107 contigs with a total genome size of 4.9 Mbp and 52% G+C content. The contigs were annotated for phylogenetic and functional analysis. Results Through the analysis, it is revealed that the genome were resistant to a number of antimicrobial drug classes including aminoglycoside, fluoroquinolone, tetracycline and phenicol. Also found in UPM 260 genome were three intact prophages (Fels-1, Gifsy-2 and one unique prophage, mEp390). The genome housed four types of restriction-modification systems (RMS) and Type I-E subtype of CRISPR-Cas system. Two metal resistance operons (mer and cop) and six pathogenicity islands (SPIs) were also discovered in UPM 260 genome. The SPIs contributed mostly to the bacterial virulence properties since 1054 CDS were reported to be homologous to the virulence factors in the database VFDB. Conclusion This study benefits us specifically in the field of genome engineering where gene-based genetic manipulations can be applied in reducing the prevalence and pathogenicity in Salmonella.

scholarly journals Functional Characterization of the Initiation Enzyme of S-Layer Glycoprotein Glycan Biosynthesis in Geobacillus stearothermophilus NRS 2004/3a

2007 ◽  
Vol 189 (7) ◽  
pp. 2590-2598 ◽  
Author(s):  
Kerstin Steiner ◽  
René Novotny ◽  
Kinnari Patel ◽  
Evgenij Vinogradov ◽  
Chris Whitfield ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Biochemical Characterization ◽  
Functional Characterization ◽  
Geobacillus Stearothermophilus ◽  
Polysaccharide Biosynthesis ◽  
Glycan Chain ◽  
Galactose Residue ◽  
Serovar Typhimurium

ABSTRACTThe glycan chain of the S-layer glycoprotein ofGeobacillus stearothermophilusNRS 2004/3a is composed of repeating units [→2)-α-l-Rhap-(1→3)-β-l-Rhap-(1→2)-α-l-Rhap-(1→], with a 2-O-methyl modification of the terminal trisaccharide at the nonreducing end of the glycan chain, a core saccharide composed of two or three α-l-rhamnose residues, and a β-d-galactose residue as a linker to the S-layer protein. In this study, we report the biochemical characterization of WsaP of the S-layer glycosylation gene cluster as a UDP-Gal:phosphoryl-polyprenol Gal-1-phosphate transferase that primes the S-layer glycoprotein glycan biosynthesis ofGeobacillus stearothermophilusNRS 2004/3a. Our results demonstrate that the enzyme transfers in vitro a galactose-1-phosphate from UDP-galactose to endogenous phosphoryl-polyprenol and that the C-terminal half of WsaP carries the galactosyltransferase function, as already observed for the UDP-Gal:phosphoryl-polyprenol Gal-1-phosphate transferase WbaP fromSalmonella enterica. To confirm the function of the enzyme, we show that WsaP is capable of reconstituting polysaccharide biosynthesis in WbaP-deficient strains ofEscherichia coliandSalmonella entericaserovar Typhimurium.

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