Limits...
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

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

License
getmorefigures.php?uid=PMC4493096&req=5

pone.0132389.g009: Structures of glycans derived from glycolipids observed in the MALDI-TOF MS spectra of Hela and NHDF cells.All glycans are deuteroreduced (DR), permethylated and shown in the form of [M+Na]+. These glycan structures and linkages are drawn based on the molecular weight, glycolipid glycan biosynthetic pathway and MS/MS data. ND, not detected. * = minor (<20%), ** = medium (20–50%), *** = major (>50%).

Mentions: MALDI-TOF data from glycans derived from the glycolipids of HeLa and NHDF cells are shown in S5 Fig. Fig 9 summarises our structural conclusions which are derived from the glycomics experiments, taking into account biosynthetic considerations. The major glycans are sialylated, and glycans terminated with uncapped Gal represent only a minor fraction. In addition, a glycan (m/z 1101) terminated with HexNAc is present in HeLa cells. Sialidase digestion of HeLa samples confirmed the α2–3 linkage of the peripheral NeuAc of GM3, GM1b and GD1a (S6 Fig).


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)

Structures of glycans derived from glycolipids observed in the MALDI-TOF MS spectra of Hela and NHDF cells.All glycans are deuteroreduced (DR), permethylated and shown in the form of [M+Na]+. These glycan structures and linkages are drawn based on the molecular weight, glycolipid 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.g009: Structures of glycans derived from glycolipids observed in the MALDI-TOF MS spectra of Hela and NHDF cells.All glycans are deuteroreduced (DR), permethylated and shown in the form of [M+Na]+. These glycan structures and linkages are drawn based on the molecular weight, glycolipid glycan biosynthetic pathway and MS/MS data. ND, not detected. * = minor (<20%), ** = medium (20–50%), *** = major (>50%).
Mentions: MALDI-TOF data from glycans derived from the glycolipids of HeLa and NHDF cells are shown in S5 Fig. Fig 9 summarises our structural conclusions which are derived from the glycomics experiments, taking into account biosynthetic considerations. The major glycans are sialylated, and glycans terminated with uncapped Gal represent only a minor fraction. In addition, a glycan (m/z 1101) terminated with HexNAc is present in HeLa cells. Sialidase digestion of HeLa samples confirmed the α2–3 linkage of the peripheral NeuAc of GM3, GM1b and GD1a (S6 Fig).

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