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Beyond Ribosomal Binding: The Increased Polarity and Aberrant Molecular Interactions of 3- epi- deoxynivalenol

View Article: PubMed Central - PubMed

ABSTRACT

Deoxynivalenol (DON) is a secondary fungal metabolite and contaminant mycotoxin that is widely detected in wheat and corn products cultivated around the world. Bio-remediation methods have been extensively studied in the past two decades and promising ways to reduce DON-associated toxicities have been reported. Bacterial epimerization of DON at the C3 carbon was recently reported to induce a significant loss in the bio-toxicity of the resulting stereoisomer (3-epi-DON) in comparison to the parental compound, DON. In an earlier study, we confirmed the diminished bio-potency of 3-epi-DON using different mammalian cell lines and mouse models and mechanistically attributed it to the reduced binding of 3-epi-DON within the ribosomal peptidyl transferase center (PTC). In the current study and by inspecting the chromatographic behavior of 3-epi-DON and its molecular interactions with a well-characterized enzyme, Fusarium graminearum Tri101 acetyltransferase, we provide the evidence that the C3 carbon epimerization of DON influences its molecular interactions beyond the abrogated PTC binding.

No MeSH data available.


Related in: MedlinePlus

Changes in 3-epi-DON concentrations in E. coli BL21(DE3) cells overexpressing Fusarium graminearum Tri101. (a) No changes in 3-epi-DON recoveries were noticed for the construct that encoded the Tri101 enzyme in comparison to the control or empty vector. The above tendency was observed in two separate experiments, each with three replications, and the different letters signify significant differences between the means according to Fisher’s Least Significant Difference (LSD) test (p < 0.005); (b) To confirm that the above cells where all over-expressing His-FgTri101, bacterial pellets were collected, lysed, and ran on SDS-PAGE gels (4%–20%) before Coomassie Blue staining. The overexpression levels of His-FgTri101 were all evident in both DON and 3-epi-DON treatments.
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toxins-08-00261-f004: Changes in 3-epi-DON concentrations in E. coli BL21(DE3) cells overexpressing Fusarium graminearum Tri101. (a) No changes in 3-epi-DON recoveries were noticed for the construct that encoded the Tri101 enzyme in comparison to the control or empty vector. The above tendency was observed in two separate experiments, each with three replications, and the different letters signify significant differences between the means according to Fisher’s Least Significant Difference (LSD) test (p < 0.005); (b) To confirm that the above cells where all over-expressing His-FgTri101, bacterial pellets were collected, lysed, and ran on SDS-PAGE gels (4%–20%) before Coomassie Blue staining. The overexpression levels of His-FgTri101 were all evident in both DON and 3-epi-DON treatments.

Mentions: On the other hand, incubation of 3-epi-DON with either E. coli BL21(DE3) cells expressing His-FgTri101 or BL21(DE3) cells with empty pET-28a(+) vector did not lead to any changes in 3-epi-DON recovery levels after methanol-extractions (Figure 4a) or to the detection of 3ADON presence in these reactions (data not shown) as a result of 3-epi-DON and FgTri101 interactions.


Beyond Ribosomal Binding: The Increased Polarity and Aberrant Molecular Interactions of 3- epi- deoxynivalenol
Changes in 3-epi-DON concentrations in E. coli BL21(DE3) cells overexpressing Fusarium graminearum Tri101. (a) No changes in 3-epi-DON recoveries were noticed for the construct that encoded the Tri101 enzyme in comparison to the control or empty vector. The above tendency was observed in two separate experiments, each with three replications, and the different letters signify significant differences between the means according to Fisher’s Least Significant Difference (LSD) test (p < 0.005); (b) To confirm that the above cells where all over-expressing His-FgTri101, bacterial pellets were collected, lysed, and ran on SDS-PAGE gels (4%–20%) before Coomassie Blue staining. The overexpression levels of His-FgTri101 were all evident in both DON and 3-epi-DON treatments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037487&req=5

toxins-08-00261-f004: Changes in 3-epi-DON concentrations in E. coli BL21(DE3) cells overexpressing Fusarium graminearum Tri101. (a) No changes in 3-epi-DON recoveries were noticed for the construct that encoded the Tri101 enzyme in comparison to the control or empty vector. The above tendency was observed in two separate experiments, each with three replications, and the different letters signify significant differences between the means according to Fisher’s Least Significant Difference (LSD) test (p < 0.005); (b) To confirm that the above cells where all over-expressing His-FgTri101, bacterial pellets were collected, lysed, and ran on SDS-PAGE gels (4%–20%) before Coomassie Blue staining. The overexpression levels of His-FgTri101 were all evident in both DON and 3-epi-DON treatments.
Mentions: On the other hand, incubation of 3-epi-DON with either E. coli BL21(DE3) cells expressing His-FgTri101 or BL21(DE3) cells with empty pET-28a(+) vector did not lead to any changes in 3-epi-DON recovery levels after methanol-extractions (Figure 4a) or to the detection of 3ADON presence in these reactions (data not shown) as a result of 3-epi-DON and FgTri101 interactions.

View Article: PubMed Central - PubMed

ABSTRACT

Deoxynivalenol (DON) is a secondary fungal metabolite and contaminant mycotoxin that is widely detected in wheat and corn products cultivated around the world. Bio-remediation methods have been extensively studied in the past two decades and promising ways to reduce DON-associated toxicities have been reported. Bacterial epimerization of DON at the C3 carbon was recently reported to induce a significant loss in the bio-toxicity of the resulting stereoisomer (3-epi-DON) in comparison to the parental compound, DON. In an earlier study, we confirmed the diminished bio-potency of 3-epi-DON using different mammalian cell lines and mouse models and mechanistically attributed it to the reduced binding of 3-epi-DON within the ribosomal peptidyl transferase center (PTC). In the current study and by inspecting the chromatographic behavior of 3-epi-DON and its molecular interactions with a well-characterized enzyme, Fusarium graminearum Tri101 acetyltransferase, we provide the evidence that the C3 carbon epimerization of DON influences its molecular interactions beyond the abrogated PTC binding.

No MeSH data available.


Related in: MedlinePlus