Dissociation of the Octameric Enolase from S. Pyogenes - One Interface Stabilizes Another

Karbassi, Farhad; Quiros, Veronica; Pancholi, Vijay; Kornblatt, Mary Judith

doi:10.1371/journal.pone.0008810

Cited by 15 publications

(30 citation statements)

References 37 publications

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“…Notably, S. gallolyticus enolase was among the abundant bacterial proteins released by the proteolytic shaving procedure (our unpublished data). These findings fit with a common mechanism for bacterial adherence to CK8 as enolase is a conserved bacterial protein with great sequence conservation between different bacterial species (22). Furthermore, enolase has already been shown to be expressed at the bacterial cell surface and to be of importance for the binding of, e.g., S. pneumoniae to airway epithelial cells (1,4,11,27,28).…”

Section: Discussionsupporting

confidence: 76%

Surface-Affinity Profiling To Identify Host-Pathogen Interactions

et al. 2011

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Proteolytic treatment of intact bacterial cells has proven to be a convenient approach for the identification of surface-exposed proteins. This class of proteins directly interacts with the outside world, for instance, during adherence to human epithelial cells. Here, we aimed to identify host receptor proteins by introducing a preincubation step in which bacterial cells were first allowed to capture human proteins from epithelial cell lysates. Using Streptococcus gallolyticus as a model bacterium, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of proteolytically released peptides yielded the identification of a selective number of human epithelial proteins that were retained by the bacterial surface. Of these potential receptors for bacterial interference, (cyto)keratin-8 (CK8) was verified as the most significant hit, and its surface localization was investigated by subcellular fractionation and confocal microscopy. Interestingly, bacterial enolase could be assigned as an interaction partner of CK8 by MS/MS analysis of cross-linked protein complexes and complementary immunoblotting experiments. As surface-exposed enolase has a proposed role in epithelial adherence of several Gram-positive pathogens, its interaction with CK8 seems to point toward a more general virulence mechanism. In conclusion, our study shows that surface-affinity profiling is a valuable tool to identify novel adhesin-receptor pairs, which advocates its application in other hybrid biological systems.The key to bacterial infection of host tissue is the establishment of a dependable connection between the bacterium and host surface structures. This is essential for the bacteria to withstand mechanical cleansing processes and to compete with other bacterial strains for microbial succession (16). After initial adherence, several pathogens can invade host cells using intracellular structures, e.g., the cytoskeleton, to sustain growth and prolong their survival times (8, 12). Ultimately, both adhesion and internalization of pathogenic bacteria will directly or indirectly (via induction of host responses) cause damage to the infected tissue. It is therefore important to fully understand the mechanisms underlying pathogenic interference so that new methods to prevent pathogenic bacteria from initiating an infectious process can be developed. In addition, knowledge about pathogen-specific interactions and subsequent responses may aid in the diagnosis of corresponding diseases.Current advances in proteomic technologies provide opportunities to compare the protein content from different biologic systems, making it possible to characterize host-pathogen interactions in a global view. Therefore, the aim of this study was to explore the use of a proteolytic shaving approach coupled to liquid chromatography-tandem mass spectrometry (LC-MS/ MS) to identify potential host proteins for bacterial interference. For this purpose, intact bacterial cells were first allowed to selectively bind host proteins from epithelial cell lysates, after wh...

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Section: Discussionsupporting

confidence: 76%

Surface-Affinity Profiling To Identify Host-Pathogen Interactions

et al. 2011

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“…Enolase has been found to exist as a homodimer in all eukaryotes and many prokaryotes (Brown et al, 1998), although octameric enolases from several bacterial species have occasionally been reported (Schurig et al, 1995;Ehinger et al, 2004;Karbassi et al, 2010). Our previous study on SS2 enolase indicates an octamer enzyme in solution by gel filtration analysis .…”

Section: Solid Evidence Of An Enolase Octamer Both In Crystal and In mentioning

confidence: 74%

“…The oligomerization state of enolase has always been an interesting issue. Most enolases are dimers in solution (Brown et al, 1998) but enolase octamers have not been rare either (Schurig et al, 1995;Ehinger et al, 2004;Karbassi et al, 2010). A previous study proposed that the length of the loop connecting H4 helix and S4 strand is possibly a key factor determining enolase octamerization (Brown et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

“…It has been proposed that most enolases exist as homodimers including those from all eukaryotes and many prokaryotes (Brown et al, 1998). In contrast, far less enolases have been demonstrated as octamers (Schurig et al, 1995;Ehinger et al, 2004;Karbassi et al, 2010), all of which are exclusively from the prokaryotic species. Therefore, the elucidation of the oligomeric state of an enolase is crucial for the characterization of the enzyme.…”

Section: Introductionmentioning

confidence: 99%

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An octamer of enolase from Streptococcus suis

Lü

et al. 2012

Protein Cell

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Enolase is a conserved cytoplasmic metalloenzyme existing universally in both eukaryotic and prokaryotic cells. The enzyme can also locate on the cell surface and bind to plasminogen, via which contributing to the mucosal surface localization of the bacterial pathogens and assisting the invasion into the host cells. The functions of the eukaryotic enzymes on the cell surface expression (including T cells, B cells, neutrophils, monocytoes, neuronal cells and epithelial cells) are not known. Streptococcus suis serotype 2 (S. suis 2, SS2) is an important zoonotic pathogen which has recently caused two large-scale outbreaks in southern China with severe streptococcal toxic shock syndrome (STSS) never seen before in human sufferers. We recently identified the SS2 enolase as an important protective antigen which could protect mice from fatal S.suis 2 infection. In this study, a 2.4-angstrom structure of the SS2 enolase is solved, revealing an octameric arrangement in the crystal. We further demonstrated that the enzyme exists exclusively as an octamer in solution via a sedimentation assay. These results indicate that the octamer is the biological unit of SS2 enolase at least in vitro and most likely in vivo as well. This is, to our knowledge, the first comprehensive characterization of the SS2 enolase octamer both structurally and biophysically, and the second octamer enolase structure in addition to that of Streptococcus pneumoniae. We also investigated the plasminogen binding property of the SS2 enzyme.

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“…Streptococcal surface enolase (SEN) is the glycolytic enzyme, α‐enolase, an octameric metalloenzyme that catalyzes the conversion of 2‐phosphoglycerate to phosphoenolpyruvate . SEN is found in the cytosol and on the surface of GAS, and possesses high binding affinity for plasmin(ogen).…”

Section: Anchorless Adhesinsmentioning

confidence: 99%

Streptococcus pyogenes adhesion and colonization

et al. 2016

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Edited by Wilhelm JustStreptococcus pyogenes (group A Streptococcus, GAS) is a human-adapted pathogen responsible for a wide spectrum of disease. GAS can cause relatively mild illnesses, such as strep throat or impetigo, and less frequent but severe life-threatening diseases such as necrotizing fasciitis and streptococcal toxic shock syndrome. GAS is an important public health problem causing significant morbidity and mortality worldwide. The main route of GAS transmission between humans is through close or direct physical contact, and particularly via respiratory droplets. The upper respiratory tract and skin are major reservoirs for GAS infections. The ability of GAS to establish an infection in the new host at these anatomical sites primarily results from two distinct physiological processes, namely bacterial adhesion and colonization. These fundamental aspects of pathogenesis rely upon a variety of GAS virulence factors, which are usually under strict transcriptional regulation. Considerable progress has been made in better understanding these initial infection steps. This review summarizes our current knowledge of the molecular mechanisms of GAS adhesion and colonization.

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Dissociation of the Octameric Enolase from S. Pyogenes - One Interface Stabilizes Another

Cited by 15 publications

References 37 publications

Surface-Affinity Profiling To Identify Host-Pathogen Interactions

Surface-Affinity Profiling To Identify Host-Pathogen Interactions

An octamer of enolase from Streptococcus suis

Streptococcus pyogenes adhesion and colonization

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