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Label-free mass spectrometry proteome quantification of human embryonic kidney cells following 24 hours of sialic acid overproduction.

Parviainen VI, Joenväärä S, Tohmola N, Renkonen R - Proteome Sci (2013)

Bottom Line: Cell surface glycoprotein sialylation is one of the most ubiquitous glycan modifications found on higher eukaryotes.Mass spectrometric analysis of cellular proteome of control and induced cells identified 1802 proteins of which 105 displayed significant changes in abundance.Functional analysis of the resulting relative changes in protein abundance revealed regulation of several cellular pathways including protein transport, metabolic and signaling pathways and remodeling of epithelial adherens junctions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Transplantation Laboratory, Haartman Institute, University of Helsinki & HUSLAB, Helsinki University Central Hospital, Helsinki, Finland. ville.parviainen@helsinki.fi.

ABSTRACT

Background: Cell surface glycoprotein sialylation is one of the most ubiquitous glycan modifications found on higher eukaryotes. The surface sialylation pattern of cells is influenced by the cellular environment but also by the Golgi sialyltransferase activity and flux of metabolites through sialic acid producing pathways. Altered cell surface sialic acid patterns have been observed in several cancers and other pathological conditions. In this experiment we examined the cellular proteomic changes that occur in human embryonic kidney cells after 24 hours of sialic acid overproduction using N-Acetylmannosamine. We utilized high resolution mass spectrometry and label free protein quantification to characterize the relative changes in protein abundance as well as multiple reaction monitoring to quantify the cellular sialic acid levels.

Results: Using N-Acetylmannosamine we were able to induce sialic acid production to almost 70-fold compared to non-induced control cells. Mass spectrometric analysis of cellular proteome of control and induced cells identified 1802 proteins of which 105 displayed significant changes in abundance. Functional analysis of the resulting relative changes in protein abundance revealed regulation of several cellular pathways including protein transport, metabolic and signaling pathways and remodeling of epithelial adherens junctions. We also identified several physically interacting co-regulated proteins in the set of changed proteins.

Conclusions: In this experiment we show that increased metabolic flux through sialic acid producing pathway affects the abundance of several protein transport, epithelial adherens junction, signaling and metabolic pathway related proteins.

No MeSH data available.


Related in: MedlinePlus

Interconnected IPA pathways. Two distinct interconnecting clusters were seen in enriched IPA pathways. One related to signaling pathways and Remodeling of adherens junctions, other related to cell cycle and translation. The connections between nodes represent shared proteins and the color intensity of the nodes reflects the increased enrichment value.
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Figure 3: Interconnected IPA pathways. Two distinct interconnecting clusters were seen in enriched IPA pathways. One related to signaling pathways and Remodeling of adherens junctions, other related to cell cycle and translation. The connections between nodes represent shared proteins and the color intensity of the nodes reflects the increased enrichment value.

Mentions: Analysis of pathways that share protein members revealed two distinct interconnected clusters (Figure 3). Larger of these networks contained several signaling networks involved in cellular movement, cell-cell contact and cytoskeletal reorganization. Most of which are also connected to Remodeling of epithelial adherens junction pathway. Second interconnected cluster contained categories on cell cycle, apoptosis and protein synthesis.


Label-free mass spectrometry proteome quantification of human embryonic kidney cells following 24 hours of sialic acid overproduction.

Parviainen VI, Joenväärä S, Tohmola N, Renkonen R - Proteome Sci (2013)

Interconnected IPA pathways. Two distinct interconnecting clusters were seen in enriched IPA pathways. One related to signaling pathways and Remodeling of adherens junctions, other related to cell cycle and translation. The connections between nodes represent shared proteins and the color intensity of the nodes reflects the increased enrichment value.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750590&req=5

Figure 3: Interconnected IPA pathways. Two distinct interconnecting clusters were seen in enriched IPA pathways. One related to signaling pathways and Remodeling of adherens junctions, other related to cell cycle and translation. The connections between nodes represent shared proteins and the color intensity of the nodes reflects the increased enrichment value.
Mentions: Analysis of pathways that share protein members revealed two distinct interconnected clusters (Figure 3). Larger of these networks contained several signaling networks involved in cellular movement, cell-cell contact and cytoskeletal reorganization. Most of which are also connected to Remodeling of epithelial adherens junction pathway. Second interconnected cluster contained categories on cell cycle, apoptosis and protein synthesis.

Bottom Line: Cell surface glycoprotein sialylation is one of the most ubiquitous glycan modifications found on higher eukaryotes.Mass spectrometric analysis of cellular proteome of control and induced cells identified 1802 proteins of which 105 displayed significant changes in abundance.Functional analysis of the resulting relative changes in protein abundance revealed regulation of several cellular pathways including protein transport, metabolic and signaling pathways and remodeling of epithelial adherens junctions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Transplantation Laboratory, Haartman Institute, University of Helsinki & HUSLAB, Helsinki University Central Hospital, Helsinki, Finland. ville.parviainen@helsinki.fi.

ABSTRACT

Background: Cell surface glycoprotein sialylation is one of the most ubiquitous glycan modifications found on higher eukaryotes. The surface sialylation pattern of cells is influenced by the cellular environment but also by the Golgi sialyltransferase activity and flux of metabolites through sialic acid producing pathways. Altered cell surface sialic acid patterns have been observed in several cancers and other pathological conditions. In this experiment we examined the cellular proteomic changes that occur in human embryonic kidney cells after 24 hours of sialic acid overproduction using N-Acetylmannosamine. We utilized high resolution mass spectrometry and label free protein quantification to characterize the relative changes in protein abundance as well as multiple reaction monitoring to quantify the cellular sialic acid levels.

Results: Using N-Acetylmannosamine we were able to induce sialic acid production to almost 70-fold compared to non-induced control cells. Mass spectrometric analysis of cellular proteome of control and induced cells identified 1802 proteins of which 105 displayed significant changes in abundance. Functional analysis of the resulting relative changes in protein abundance revealed regulation of several cellular pathways including protein transport, metabolic and signaling pathways and remodeling of epithelial adherens junctions. We also identified several physically interacting co-regulated proteins in the set of changed proteins.

Conclusions: In this experiment we show that increased metabolic flux through sialic acid producing pathway affects the abundance of several protein transport, epithelial adherens junction, signaling and metabolic pathway related proteins.

No MeSH data available.


Related in: MedlinePlus