Cloning of the Tannerella forsythensis (Bacteroides forsythus) siaHI gene and purification of the sialidase enzyme

Ishikura, Hiroaki; Arakawa, Shin’ichi; Nakajima, Takuma; Tsuchida, Nobuo; Ishikawa, Isao

doi:10.1099/jmm.0.05349-0

Cited by 44 publications

(43 citation statements)

References 25 publications

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“…Further, the post-translational addition of carbohydrates to the gingipains is highly variable, thus implicating a role for multiple factors in this process. In this study, the VimA protein was also observed to interact with the b-lactamase and sialidase proteins,which are known to play a role in sugar metabolism (Ishikura et al, 2003;Smayevsky et al, 2001;Brook, 1989;Hedberg & Nord, 1996;Sechi et al, 2004). Although sialylation can be involved in protein maturation, its role, if any, in gingipain biogenesis is unknown.…”

Section: Discussionmentioning

confidence: 88%

VimA is part of the maturation pathway for the major gingipains of Porphyromonas gingivalis W83

et al. 2006

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The authors have shown previously that the vimA gene, which is part of the bcp-recA-vimA operon, plays an important role in protease activation in Porphyromonas gingivalis. The gingipain RgpB proenzyme is secreted in the vimA-defective mutant P. gingivalis FLL92. An important question that is raised is whether the vimA gene product could directly interact with the proteases for their activation or regulate a pathway responsible for protease activation. To further study the mechanism(s) of VimA-dependent protease activation, the vimA gene product was further characterized. A 39 kDa protein consistent with the size of the predicted VimA protein was purified. In protein–protein interaction studies, the VimA protein was shown to interact with gingipains RgpA, RgpB and Kgp. Immune sera from mice immunized with P. gingivalis immunoreacted with the purified VimA protein. Taken together, these data suggest an interaction of VimA with the gingipains and further confirm the role of this protein in their regulation or maturation.

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Section: Discussionmentioning

confidence: 88%

VimA is part of the maturation pathway for the major gingipains of Porphyromonas gingivalis W83

et al. 2006

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“…The previously described SiaHI protein of T. forsythia (19) does not account for all of the sialidase activity of this putative periodontal pathogen. This bacterium has an orthologue of B. fragilis NanH, TF0035 (renamed NanH), which has sialidase activity when expressed in E. coli.…”

Section: Discussionmentioning

confidence: 99%

“…Sialic acids were eluted from the column by using a linear gradient of sodium acetate (20 to 150 mM over 40 min) in 100 mM sodium hydroxide at a flow rate of 1 ml/min. N-Acetylneuraminic acid and N-glycolylneuraminic acid concentrations in assays were determined by comparison of peak areas with those of the authentic sialic acids (Sigma) as standards over a concentration range of 0 to 50 M. (19), it is not apparent whether this alone is sufficient to account for all of the sialidase activity in this bacterium. Sialidase activity not directly attributable to SiaHI was observed upon nondenaturing gel electrophoresis of T. forsythia proteins (19).…”

Section: Methodsmentioning

confidence: 99%

“…N-Acetylneuraminic acid and N-glycolylneuraminic acid concentrations in assays were determined by comparison of peak areas with those of the authentic sialic acids (Sigma) as standards over a concentration range of 0 to 50 M. (19), it is not apparent whether this alone is sufficient to account for all of the sialidase activity in this bacterium. Sialidase activity not directly attributable to SiaHI was observed upon nondenaturing gel electrophoresis of T. forsythia proteins (19). Also, SiaHI is an unusual sialidase since it does not contain motifs associated with other characterized sialidases (31).…”

Section: Methodsmentioning

confidence: 99%

“…Although a T. forsythia sialidase gene, siaHI, has been reported (19), it is unclear whether this alone is responsible for sialidase activity in T. forsythia. A sialidase unrelated to T. forsythia SiaHI has been described in the related bacterium, Bacteroides fragilis (49), and it may be important for nutrient acquisition, supporting the growth of this bacterium in vivo (14).…”

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An Orthologue of Bacteroides fragilis NanH Is the Principal Sialidase in Tannerella forsythia

Thompson¹,

Homer²,

Rao³

et al. 2009

J Bacteriol

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Sialidase activity is a putative virulence factor of the anaerobic periodontal pathogen Tannerella forsythia, but it is uncertain which genes encode this activity. Characterization of a putative sialidase, SiaHI, by others, indicated that this protein alone may not be responsible for all of the sialidase activity. We describe a second sialidase in T. forsythia (TF0035), an orthologue of Bacteroides fragilis NanH, and its expression in Escherichia coli. Sialidase activity of the expressed NanH was confirmed by using 2-(4-methylumbelliferyl)-␣-D-N-acetylneuraminic acid as a substrate. Biochemical characterization of the recombinant T. forsythia NanH indicated that it was active over a broad pH range, with optimum activity at pH 5.5. This enzyme has high affinity for 2-(4-methylumbelliferyl)-␣-D-N-acetylneuraminic acid (K m of 32.9 ؎ 10.3 M) and rapidly releases 4-methylumbelliferone (V max of 170.8 ؎ 11.8 nmol of 4-methylumbelliferone min ؊1 mg of protein ؊1 ). E. coli lysates containing recombinant T. forsythia NanH cleave sialic acid from a range of substrates, with a preference for ␣2-3 glycosidic linkages. The genes adjacent to nanH encode proteins apparently involved in the metabolism of sialic acid, indicating that the NanH sialidase is likely to be involved in nutrient acquisition.

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Tannerella

Sakamoto¹,

Benno²

2015

Bergey's Manual of Systematics of Archaea and Bacteria

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Tan.ne.rel'la. L. dim. suffix ‐ ella ; N.L. fem. dim. n. Tannerella named after the American microbiologist Anne C. R. Tanner, for her contributions to research on periodontal disease. Bacteroidetes / Bacteroidia / Bacteroidales / Porphyromonadaceae / Tannerella Fusiform cells , generally 0.3–0.5 × 1–30 µm. Gram‐negative. Nonmotile. Obligately anaerobic . N ‐ acetylmuramic acid ( NAM ) is required for growth (some strains do not require NAM). Growth is inhibited in the presence of 20 % bile . The major end products are acetic acid, butyric acid, isovaleric acid, propionic acid, and phenylacetic acid; smaller amounts of isobutyric acid and succinic acid may be produced. Esculin is hydrolyzed. Indole variable. Trypsin activity is positive . Glucose‐6‐phosphate dehydrogenase (G6PDH), 6‐phosphogluconate dehydrogenase (6PGDH), malate dehydrogenase, and glutamate dehydrogenase are present. The principal respiratory quinones are menaquinones MK‐10 and MK‐11. Both nonhydroxylated and 3‐hydroxylated long‐chain fatty acids are present. The nonhydroxylated acids are predominantly of the saturated straight‐chain and anteiso methyl‐branched‐chain types. The ratio of C 15 : 0 anteiso to C 15: 0 iso is high (> 20 ). Isolated originally and principally from human oral cavity . Tannerella forsythia has been associated with periodontal disease , root canal infections , and peri‐implantitis . DNA G + C content ( mol %): 44–48. Type species : Tannerella forsythia corrig. (Tanner, Listgarten, Ebersole and Strezempko, 1986) Sakamoto, Suzuki, Umeda, Ishikawa and Benno 2002, 848 VP ( Bacteroides forsythus Tanner, Listgarten, Ebersole and Strezempko, 1986, 216).

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Cloning of the Tannerella forsythensis (Bacteroides forsythus) siaHI gene and purification of the sialidase enzyme

Cited by 44 publications

References 25 publications

VimA is part of the maturation pathway for the major gingipains of Porphyromonas gingivalis W83

VimA is part of the maturation pathway for the major gingipains of Porphyromonas gingivalis W83

An Orthologue of Bacteroides fragilis NanH Is the Principal Sialidase in Tannerella forsythia

Tannerella

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