The sialidase gene from Clostridium septicum: cloning, sequencing, expression in Escherichia coli and identification of conserved sequences in sialidases and other proteins

Rothe, Bernd; Roggentin, Peter; Schauer, Roland

doi:10.1007/bf00273603

Cited by 65 publications

(33 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Comparison of the NanB sequence with that of NanA revealed negligible homology. However, comparison with other sequences deposited with GenBank indicated a limited degree of amino acid homology (30% identity over 169 residues) with the sialidase of Clostridium septicum (30). The region of greatest similarity (51% identity and 86% similarity over 51 residues) includes an R-I-P motif that is found in the active site of other bacterial neuraminidases (29).…”

Section: Resultsmentioning

confidence: 98%

Cloning and characterization of nanB, a second Streptococcus pneumoniae neuraminidase gene, and purification of the NanB enzyme from recombinant Escherichia coli

1996

View full text Add to dashboard Cite

Streptococcus pneumoniae is believed to produce more than one form of neuraminidase, but there has been uncertainty as to whether this is due to posttranslational modification of a single gene product or the existence of more than one neuraminidase-encoding gene. Only one stable pneumococcal neuraminidase gene (designated nanA) has been described. In the present study, we isolated and characterized a second neuraminidase gene (designated nanB), which is located close to nanA on the pneumococcal chromosome (approximately 4.5 kb downstream). nanB was located on an operon separate from that of nanA, which includes at least five other open reading frames. NanB has a predicted size of 74.5 kDa after cleavage of a 29-amino-acid signal peptide. There was negligible amino acid homology between NanA and NanB, but NanB did exhibit limited homology with the sialidase of Clostridium septicum. NanB was purified from recombinant Escherichia coli and found to have a pH optimum of 4.5, compared with 6.5 to 7.0 for NanA. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis suggested that NanB has a molecular size of approximately 65 kDa. The discrepancy between this estimate and the size predicted from the nucleotide sequence is most likely a consequence of C-terminal processing or anomalous electrophoretic behavior.

show abstract

Section: Resultsmentioning

confidence: 98%

Cloning and characterization of nanB, a second Streptococcus pneumoniae neuraminidase gene, and purification of the NanB enzyme from recombinant Escherichia coli

1996

View full text Add to dashboard Cite

show abstract

“…Viral neuraminidase inhibitors have been very useful in the prevention and treatment of influenza, targeting similar high-risk patient populations. The PA2794 neuraminidase shares many conserved elements and folds in the manner predicted for other microbial neuraminidases (19,55). A preliminary analysis of PA2794 crystal structure indicates that the enzyme shares the same sialic acid binding fold as does the influenza enzyme, but otherwise has little homology (L. Tong and Y.-S. Hsiao, unpublished observations).…”

Section: Discussionmentioning

confidence: 95%

Bacterial neuraminidase facilitates mucosal infection by participating in biofilm production

Soong¹

2006

Journal of Clinical Investigation

179

117

View full text Add to dashboard Cite

Many respiratory pathogens, including Hemophilus influenzae, Streptococcus pneumoniae, and Pseudomonas aeruginosa, express neuraminidases that can cleave α2,3-linked sialic acids from glycoconjugates. As mucosal surfaces are heavily sialylated, neuraminidases have been thought to modify epithelial cells by exposing potential bacterial receptors. However, in contrast to neuraminidase produced by the influenza virus, a role for bacterial neuraminidase in pathogenesis has not yet been clearly established. We constructed a mutant of P. aeruginosa PAO1 by deleting the PA2794 neuraminidase locus (D2794) and tested its virulence and immunostimulatory capabilities in a mouse model of infection. Although fully virulent when introduced i.p., the D2794 mutant was unable to establish respiratory infection by i.n. inoculation. The inability to colonize the respiratory tract correlated with diminished production of biofilm, as assessed by scanning electron microscopy and in vitro assays. The importance of neuraminidase in biofilm production was further demonstrated by showing that viral neuraminidase inhibitors in clinical use blocked P. aeruginosa biofilm production in vitro as well. The P. aeruginosa neuraminidase has a key role in the initial stages of pulmonary infection by targeting bacterial glycoconjugates and contributing to the formation of biofilm. Inhibiting bacterial neuraminidases could provide a novel mechanism to prevent bacterial pneumonia.

show abstract

“…The nanI gene (Locus CPE 0725) encodes a 694-amino acid protein known as the "large sialidase enzyme" (49). The nanJ gene (Locus CPE 0553) encodes a 1173-amino acid protein not yet characterized, which displays a 51% sequence similarity to the 111-kDa sialidase from Clostridium septicum (50). Sialidase activity is detectable in culture supernatants during exponential growth of C. perfringens, and in vivo it is detectable in the bloodstream of experimental animals or patients with gas gangrene in early stages of the disease (51,52).…”

Section: Discussionmentioning

confidence: 99%

A Cellular Deficiency of Gangliosides Causes Hypersensitivity to Clostridium perfringens Phospholipase C

Flores-Dı́az

Alape-Girón

Clark

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Clostridium perfringens phospholipase C (Cp-PLC), also called ␣-toxin, is the major virulence factor in the pathogenesis of gas gangrene. Previously, a cellular UDP-Glc deficiency was related with a hypersensitivity to the cytotoxic effect of Cp-PLC. Because UDP-Glc is required in the synthesis of proteoglycans, N-linked glycoproteins, and glycosphingolipids, the role of these glycoconjugates in the cellular sensitivity to Cp-PLC was studied. The cellular sensitivity to Cp-PLC was significantly enhanced by glycosphingolipid synthesis inhibitors, and a mutant cell line deficient in gangliosides was found to be hypersensitive to Cp-PLC. Gangliosides protected hypersensitive cells from the cytotoxic effect of Cp-PLC and prevented its membrane-disrupting effect on artificial membranes. Removal of sialic acids by C. perfringens sialidase increases the sensitivity of cultured cells to Cp-PLC and intramuscular co-injection of C. perfringens sialidase, and Cp-PLC in mice potentiates the myotoxic effect of the latter. This work demonstrated that a reduction in gangliosides renders cells more susceptible to the membrane damage caused by Cp-PLC and revealed a previously unrecognized synergism between Cp-PLC and C. perfringens sialidase, providing new insights toward understanding the pathogenesis of clostridial myonecrosis.

show abstract

The sialidase gene from Clostridium septicum: cloning, sequencing, expression in Escherichia coli and identification of conserved sequences in sialidases and other proteins

Cited by 65 publications

References 31 publications

Cloning and characterization of nanB, a second Streptococcus pneumoniae neuraminidase gene, and purification of the NanB enzyme from recombinant Escherichia coli

Cloning and characterization of nanB, a second Streptococcus pneumoniae neuraminidase gene, and purification of the NanB enzyme from recombinant Escherichia coli

Bacterial neuraminidase facilitates mucosal infection by participating in biofilm production

A Cellular Deficiency of Gangliosides Causes Hypersensitivity to Clostridium perfringens Phospholipase C

Contact Info

Product

Resources

About