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The Cytotoxicity of Elderberry Ribosome-Inactivating Proteins Is Not Solely Determined by Their Protein Translation Inhibition Activity.

Shang C, Chen Q, Dell A, Haslam SM, De Vos WH, Van Damme EJ - PLoS ONE (2015)

Bottom Line: In this study, we compared the in vitro and intracellular activity of several S. nigra (elderberry) RIPs and non-RIP lectins.Despite the fact that the bulk of the RIPs accumulated in the lysosomes and partly in the Golgi apparatus, we could demonstrate effective inhibition of protein synthesis in cellula.Our data suggest that one of these pathways involves the induction of autophagy.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.

ABSTRACT
Although the protein translation inhibition activity of ribosome inactivating proteins (RIPs) is well documented, little is known about the contribution of the lectin chain to the biological activity of these proteins. In this study, we compared the in vitro and intracellular activity of several S. nigra (elderberry) RIPs and non-RIP lectins. Our data demonstrate that RIPs from elderberry are much more toxic to HeLa cells than to primary fibroblasts. Differences in the cytotoxicity between the elderberry proteins correlated with differences in glycan specificity of their lectin domain, cellular uptake efficiency and intracellular destination. Despite the fact that the bulk of the RIPs accumulated in the lysosomes and partly in the Golgi apparatus, we could demonstrate effective inhibition of protein synthesis in cellula. As we also observed cytotoxicity for non-RIP lectins, it is clear that the lectin chain triggers additional pathways heralding cell death. Our data suggest that one of these pathways involves the induction of autophagy.

No MeSH data available.


Related in: MedlinePlus

Effect of the S. nigra RIPs and lectins on protein synthesis in a cell-free translation assay.(A) Dose response curves of luciferase synthesis in treatments with SNA-I (non-reduced and reduced) and lectins (SNA-II, SNA-IV). (B and C) Dose response curves of luciferase synthesis in the treatments with non-reduced and reduced RIP for SNA-V and SNLRP, respectively. (D) IC50 values for the RIPs and lectins.
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pone.0132389.g001: Effect of the S. nigra RIPs and lectins on protein synthesis in a cell-free translation assay.(A) Dose response curves of luciferase synthesis in treatments with SNA-I (non-reduced and reduced) and lectins (SNA-II, SNA-IV). (B and C) Dose response curves of luciferase synthesis in the treatments with non-reduced and reduced RIP for SNA-V and SNLRP, respectively. (D) IC50 values for the RIPs and lectins.

Mentions: In an in vitro cell-free system, all non-reduced and reduced RIPs (SNA-I, SNA-V and SNLRP) from S. nigra clearly showed a concentration-dependent translation-inhibiting activity, albeit with strongly varying potency (Fig 1A and 1B) (S1 Fig). Compared to SNA-I (IC50 = 5.48 nM), the IC50 values for S. nigra RIPs SNA-V and SNLRP were significantly lower (25 to 50 fold) (Fig 1D), indicating that their activity was proportionally higher. The lectins SNA-II and SNA-IV, which are merely composed of the lectin chain, also started interfering with the translation process in the dose range of SNA-I (Fig 1A). The protein synthesis inhibition activity was not enhanced by DTT-mediated reduction of the proteins since the IC50 values for the non-reduced and reduced SNA-I, SNA-V and SNLRP are very similar (Fig 1).


The Cytotoxicity of Elderberry Ribosome-Inactivating Proteins Is Not Solely Determined by Their Protein Translation Inhibition Activity.

Shang C, Chen Q, Dell A, Haslam SM, De Vos WH, Van Damme EJ - PLoS ONE (2015)

Effect of the S. nigra RIPs and lectins on protein synthesis in a cell-free translation assay.(A) Dose response curves of luciferase synthesis in treatments with SNA-I (non-reduced and reduced) and lectins (SNA-II, SNA-IV). (B and C) Dose response curves of luciferase synthesis in the treatments with non-reduced and reduced RIP for SNA-V and SNLRP, respectively. (D) IC50 values for the RIPs and lectins.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493096&req=5

pone.0132389.g001: Effect of the S. nigra RIPs and lectins on protein synthesis in a cell-free translation assay.(A) Dose response curves of luciferase synthesis in treatments with SNA-I (non-reduced and reduced) and lectins (SNA-II, SNA-IV). (B and C) Dose response curves of luciferase synthesis in the treatments with non-reduced and reduced RIP for SNA-V and SNLRP, respectively. (D) IC50 values for the RIPs and lectins.
Mentions: In an in vitro cell-free system, all non-reduced and reduced RIPs (SNA-I, SNA-V and SNLRP) from S. nigra clearly showed a concentration-dependent translation-inhibiting activity, albeit with strongly varying potency (Fig 1A and 1B) (S1 Fig). Compared to SNA-I (IC50 = 5.48 nM), the IC50 values for S. nigra RIPs SNA-V and SNLRP were significantly lower (25 to 50 fold) (Fig 1D), indicating that their activity was proportionally higher. The lectins SNA-II and SNA-IV, which are merely composed of the lectin chain, also started interfering with the translation process in the dose range of SNA-I (Fig 1A). The protein synthesis inhibition activity was not enhanced by DTT-mediated reduction of the proteins since the IC50 values for the non-reduced and reduced SNA-I, SNA-V and SNLRP are very similar (Fig 1).

Bottom Line: In this study, we compared the in vitro and intracellular activity of several S. nigra (elderberry) RIPs and non-RIP lectins.Despite the fact that the bulk of the RIPs accumulated in the lysosomes and partly in the Golgi apparatus, we could demonstrate effective inhibition of protein synthesis in cellula.Our data suggest that one of these pathways involves the induction of autophagy.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.

ABSTRACT
Although the protein translation inhibition activity of ribosome inactivating proteins (RIPs) is well documented, little is known about the contribution of the lectin chain to the biological activity of these proteins. In this study, we compared the in vitro and intracellular activity of several S. nigra (elderberry) RIPs and non-RIP lectins. Our data demonstrate that RIPs from elderberry are much more toxic to HeLa cells than to primary fibroblasts. Differences in the cytotoxicity between the elderberry proteins correlated with differences in glycan specificity of their lectin domain, cellular uptake efficiency and intracellular destination. Despite the fact that the bulk of the RIPs accumulated in the lysosomes and partly in the Golgi apparatus, we could demonstrate effective inhibition of protein synthesis in cellula. As we also observed cytotoxicity for non-RIP lectins, it is clear that the lectin chain triggers additional pathways heralding cell death. Our data suggest that one of these pathways involves the induction of autophagy.

No MeSH data available.


Related in: MedlinePlus