Bacterial elongation factors EF-Tu, their mutants, chimeric forms, and domains: Isolation and purification

Jonák, Jiřı́

doi:10.1016/j.jchromb.2006.11.053

Cited by 16 publications

(14 citation statements)

References 92 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…EF‐Tu is a guanine nucleotide‐binding protein that plays a central role in protein synthesis. EF‐Tu is composed of three domains and the N‐terminally located domain 1 contains the guanine nucleotide‐binding site and is also called the catalytic or the G‐domain (for a review, see Sanderova et al , 2004; Sanderova & Jonak, 2005; Jonak, 2007 and references therein). Domain 2 and domain 3 are noncatalytic domains found to modulate properties and activities of domain 1.…”

Section: Resultsmentioning

confidence: 99%

“…Pneumococci were cultured in Todd–Hewitt broth (Roth, Karlsruhe, Germany) supplemented with 0.5% yeast extract (THY). Neisseria meningitidis group B clinical Norwegian isolate 44/76 (B:15: P1.7,16) and group B clinical New Zealand isolate NZ98/254 (B:4:P1.7b,4) were cultivated in Frantz' medium with shaking E. coli B, Bacillus stearothermophilus CCM 2184 and Bacillus subtilis 168 used for isolation of EF‐Tu are described in Jonak (2007) and references therein. The other bacteria (besides Lactococcus lactis IL 1403) that were used in this study have been described previously (Kolberg et al , 1997b).…”

Section: Methodsmentioning

confidence: 99%

“…EF‐Tu from E. coli , B. stearothermophilus and B. subtilis were prepared as described in Jonak (2007), and references therein. Recombinant G‐domains of E. coli EF‐Tu and B. stearothermophilus EF‐Tu were prepared as described in Tomincova et al (2002), Sanderova et al (2004), and Jonak (2007).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

The surface-associated elongation factor Tu is concealed for antibody binding on viable pneumococci and meningococci

Kolberg

Hammerschmidt

Frank

et al. 2008

FEMS Immunol Med Microbiol

View full text Add to dashboard Cite

Proteome analyses revealed that elongation factor-Tu (EF-Tu) is associated with cytoplasmic membranes of Gram-positive bacteria and outer membranes of Gram-negative bacteria. It is still debatable whether EF-Tu is located on the external side or the internal side of the membranes. Here, we have generated two new monoclonal antibodies (mAbs) and polyclonal rabbit antibodies against pneumococcal EF-Tu. These antibodies were used to investigate the amount of surface-exposed EF-Tu on viable bacteria using a flow cytometric analysis. The control antibodies recognizing the pneumococcal surface protein A and phosphorylcholine showed a significant binding to viable pneumococci. In contrast, anti-EF-Tu antibodies did not recognize pneumococcal EF-Tu. However, heat killing of pneumococci lacking capsular polysaccharides resulted in specific antibody binding to EF-Tu and, moreover, increased the exposure of recognized phosphorylcholine epitopes. Similarly, our EF-Tu-specific antibodies did not recognize EF-Tu of viable Neisseria meningitidis. However, pretreatment of meningococci with ethanol resulted in specific antibody binding to EF-Tu on outer membranes. Importantly, these treatments did not destroy the membrane integrity as analysed with control mAbs directed against cytoplasmic proteins. In conclusion, our flow cytrometric assays emphasize the importance of using viable bacteria and not heat-killed or ethanol-treated bacteria for surface-localization experiments of proteins, because these treatments modulate the cytoplasmic and outer membranes of bacteria and the binding results may not reflect the situation under physiological conditions.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The surface-associated elongation factor Tu is concealed for antibody binding on viable pneumococci and meningococci

Kolberg

Hammerschmidt

Frank

et al. 2008

FEMS Immunol Med Microbiol

View full text Add to dashboard Cite

show abstract

“…Tuf is a relatively conserved protein and has been identified from other pathogenic as well as non pathogenic species, including Gram positive and Gram negative bacteria, like L. monocytogenes and E. coli, fungi like C. albicans and Saccharomyces cerevisiae, as well as viruses, multicellular organisms and vertebrates (Archambaud et al, 2005;Crowe et al, 2003;Gross and Kinzy, 2005;Jacobson and Rosenbusch, 1976;Jonak, 2007;Li et al, 2013). It will be of interest to define if Tuf expressed by these other pathogenic species share these immune evasion properties.…”

Section: Discussionmentioning

confidence: 99%

Tuf of Streptococcus pneumoniae is a surface displayed human complement regulator binding protein

Mohan

Hertweck

Dudda

et al. 2014

Molecular Immunology

View full text Add to dashboard Cite

“…60% of model animals immunized by the rEF-Tu protein with propolis as adjuvant were protected after injection of SS2 as a virulent pathogen. Many studies have demonstrated that EF-Tu is a conserved-multifunctional bacterial protein and is an important part of the bacterial membrane cytoskeleton (Mayer 2003, Jonak 2007. A recent study indicate that EF-Tu may play a formerly under-appreciated role as a bacterial virulence factor and have an evident effect to promote early colonization of host cells of Pseudomonas aeruginosa (Barbier et al 2013).…”

Section: Immunoprotection Of Ref-tu Protein In Rabbitsmentioning

confidence: 99%

Expression and immunological evaluation of elongation factor Tu of Streptococcus suis serotype 2

Xia¹,

Wang²,

Cheng³

et al. 2017

Polish Journal of Veterinary Sciences

View full text Add to dashboard Cite

Streptococcus suis serotype 2 (SS2) is considered as a major pathogen that causes sepsis and meningitis in piglets and humans, but knowledge of its antigenic proteins remains limited so far. The surface-related proteins of pathogens often play significant roles in bacterium-host interactions and infection. Here, we obtained the elongation factor Tu (EF-Tu) gene of Streptococcus suis and constructed the recombinant expression plasmid successfully. The target recombinant plasmid was then expressed in Escherichia coli and the immuno-protection of the recombinant protein was subsequently evaluated as well. The EF-Tu gene of Streptococcus suis is 1197 bp in length, encodes 398 amino acids. The target recombinant EF-Tu (rEF-Tu) protein can recognize the antiserum of Streptococcus suis and can provoke obvious humoral immune responses in rabbits and conferred protection to rabbits against Streptococcus suis ear-vein challenge, implying that the EF-Tu may be used as an attractive candidate antigen for a component of subunit vaccine.

show abstract

Bacterial elongation factors EF-Tu, their mutants, chimeric forms, and domains: Isolation and purification

Cited by 16 publications

References 92 publications

The surface-associated elongation factor Tu is concealed for antibody binding on viable pneumococci and meningococci

The surface-associated elongation factor Tu is concealed for antibody binding on viable pneumococci and meningococci

Tuf of Streptococcus pneumoniae is a surface displayed human complement regulator binding protein

Expression and immunological evaluation of elongation factor Tu of Streptococcus suis serotype 2

Contact Info

Product

Resources

About