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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

O-glycan structures observed in the MALDI-TOF MS spectra of Hela and NHDF glycoproteins.All glycans are permethylated and shown in the form of [M+Na]+. Glycan structures and linkages are drawn based on the molecular weight, O-glycan biosynthetic pathway and MS/MS data. ND, not detected. * = minor (<20%), ** = medium (20–50%), *** = major (>50%).
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pone.0132389.g008: O-glycan structures observed in the MALDI-TOF MS spectra of Hela and NHDF glycoproteins.All glycans are permethylated and shown in the form of [M+Na]+. Glycan structures and linkages are drawn based on the molecular weight, O-glycan biosynthetic pathway and MS/MS data. ND, not detected. * = minor (<20%), ** = medium (20–50%), *** = major (>50%).

Mentions: MALDI-TOF spectra of O-glycans from HeLa and NHDF cells are shown in S4 Fig The profile demonstrated the presence of both core 1 and core 2 structures in NHDF cells whilst in HeLa cells the majority of the glycan structures are of the core 1 type, although minor core 2 glycans were observed after sialidase digestion (Fig 8). In both cell types the O-glycans are either sialylated with NeuAc or terminated with uncapped Gal. Comparison of the abundances of the O-glycans (Fig 8) indicated that the glycans terminated with Gal are more prominent in NHDF cells than in HeLa cells, while the relative abundance of the glycans terminated with the α2–6 linked NeuAc (NeuAc(α2–6)GalNAc) is higher in HeLa cells.


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)

O-glycan structures observed in the MALDI-TOF MS spectra of Hela and NHDF glycoproteins.All glycans are permethylated and shown in the form of [M+Na]+. Glycan structures and linkages are drawn based on the molecular weight, O-glycan biosynthetic pathway and MS/MS data. ND, not detected. * = minor (<20%), ** = medium (20–50%), *** = major (>50%).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132389.g008: O-glycan structures observed in the MALDI-TOF MS spectra of Hela and NHDF glycoproteins.All glycans are permethylated and shown in the form of [M+Na]+. Glycan structures and linkages are drawn based on the molecular weight, O-glycan biosynthetic pathway and MS/MS data. ND, not detected. * = minor (<20%), ** = medium (20–50%), *** = major (>50%).
Mentions: MALDI-TOF spectra of O-glycans from HeLa and NHDF cells are shown in S4 Fig The profile demonstrated the presence of both core 1 and core 2 structures in NHDF cells whilst in HeLa cells the majority of the glycan structures are of the core 1 type, although minor core 2 glycans were observed after sialidase digestion (Fig 8). In both cell types the O-glycans are either sialylated with NeuAc or terminated with uncapped Gal. Comparison of the abundances of the O-glycans (Fig 8) indicated that the glycans terminated with Gal are more prominent in NHDF cells than in HeLa cells, while the relative abundance of the glycans terminated with the α2–6 linked NeuAc (NeuAc(α2–6)GalNAc) is higher in HeLa cells.

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