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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

Physical interactions. Nine different physically interacting clusters were identified between 40 of the changed proteins. Functional similarities (grey boxes) between interacting proteins were found in proteasomic, ribosomal and spliceosomic proteins. Blue edges represent physical interactions between proteins. Green indicates down-regulation, red up-regulation; more intense color indicates larger change in abundance.
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Figure 4: Physical interactions. Nine different physically interacting clusters were identified between 40 of the changed proteins. Functional similarities (grey boxes) between interacting proteins were found in proteasomic, ribosomal and spliceosomic proteins. Blue edges represent physical interactions between proteins. Green indicates down-regulation, red up-regulation; more intense color indicates larger change in abundance.

Mentions: Analysis of the protein-protein interactions between the set of 105 proteins was done by downloading all the known interactions from PINA database [25,26]. This network of 2421 proteins and 4539 interactions was filtered to include only interactions between the changed proteins. 47 interactions were retained between 40 of the 105 proteins (Figure 4).


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)

Physical interactions. Nine different physically interacting clusters were identified between 40 of the changed proteins. Functional similarities (grey boxes) between interacting proteins were found in proteasomic, ribosomal and spliceosomic proteins. Blue edges represent physical interactions between proteins. Green indicates down-regulation, red up-regulation; more intense color indicates larger change in abundance.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Physical interactions. Nine different physically interacting clusters were identified between 40 of the changed proteins. Functional similarities (grey boxes) between interacting proteins were found in proteasomic, ribosomal and spliceosomic proteins. Blue edges represent physical interactions between proteins. Green indicates down-regulation, red up-regulation; more intense color indicates larger change in abundance.
Mentions: Analysis of the protein-protein interactions between the set of 105 proteins was done by downloading all the known interactions from PINA database [25,26]. This network of 2421 proteins and 4539 interactions was filtered to include only interactions between the changed proteins. 47 interactions were retained between 40 of the 105 proteins (Figure 4).

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