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The proteins of Fusobacterium spp. involved in hydrogen sulfide production from L-cysteine

Artikel i vetenskaplig tidskrift
Författare Amina Basic
M. Blomqvist
Gunnar Dahlén
G. Svensater
Publicerad i Bmc Microbiology
Volym 17
Nummer/häfte 61
ISSN 1471-2180
Publiceringsår 2017
Publicerad vid Institutionen för odontologi, sektion 3
Språk en
Länkar doi.org/10.1186/s12866-017-0967-9
Ämnesord Periodontitis, Hydrogen sulfide, Fusobacterium spp., Enzymes, Bismuth sulfide, Proteomics, 2D gel, nucleatum atcc 25586, c-s lyase, methyl mercaptan, periodontal pockets, identification, desulfhydrase, glutathione, methionine, Microbiology
Ämneskategorier Parodontologi, Mikrobiologi inom det medicinska området, Odontologi

Sammanfattning

Background: Hydrogen sulfide (H2S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease. We studied the H2S-producing protein expression of bacterial strains associated with periodontal disease. Further, we examined the effect of a cysteine-rich growth environment on the synthesis of intracellular enzymes in F. nucleatum polymorphum ATCC 10953. The proteins were subjected to one-dimensional (1DE) and two-dimensional (2DE) gel electrophoresis An in-gel activity assay was used to detect the H2S-producing enzymes; Sulfide from H2S, produced by the enzymes in the gel, reacted with bismuth forming bismuth sulfide, illustrated as brown bands (1D) or spots (2D) in the gel. The discovered proteins were identified with liquid chromatography - tandem mass spectrometry (LC-MS/MS). Results: Cysteine synthase and proteins involved in the production of the coenzyme pyridoxal 5'phosphate (that catalyzes the production of H2S) were frequently found among the discovered enzymes. Interestingly, a higher expression of H2S-producing enzymes was detected from bacteria incubated without cysteine prior to the experiment. Conclusions: Numerous enzymes, identified as cysteine synthase, were involved in the production of H2S from cysteine and the expression varied among Fusobacterium spp. and strains. No enzymes were detected with the in-gel activity assay among the other periodontitis-associated bacteria tested. The expression of the H2S-producing enzymes was dependent on environmental conditions such as cysteine concentration and pH but less dependent on the presence of serum and hemin.

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