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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

Chemical structures of ManNAc and Neu5Ac. Chemical structure representation of N-Acetylmannosamine and N-Acetylneuraminic acid.
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Figure 1: Chemical structures of ManNAc and Neu5Ac. Chemical structure representation of N-Acetylmannosamine and N-Acetylneuraminic acid.

Mentions: Sialic acids are one of the most common terminal monosaccharides found on cell surface glycans of mammals and other higher eukaryotes. Due to its ubiquity and properties sialic acids are involved in many biological functions ranging from early fetal development, cellular recognition and adhesion processes to protein half-life and utilization by influenza virus in entry to cell [1]. The term sialic acid covers more than 50 different glycan structures with a common nine carbon structural backbone and a carboxylic acid group at carbon one. The most common sialic acid structure variant and main metabolic precursor of most other sialic acids is N-Acetylneuraminic acid, Neu5Ac (Figure 1). Cellular Neu5Ac is produced by recycling it from surface glycoproteins using salvage pathway [2] or by de-novo biosynthesis route from other metabolic precursors [3]. The end product of the De novo pathway is activated in the nucleus by N-acylneuraminate cytidylyltransferase to produce CMP-Neu5Ac that can be utilized by the Golgi glycosyltransferases in biosynthesis of glycoconjugates.


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)

Chemical structures of ManNAc and Neu5Ac. Chemical structure representation of N-Acetylmannosamine and N-Acetylneuraminic acid.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Chemical structures of ManNAc and Neu5Ac. Chemical structure representation of N-Acetylmannosamine and N-Acetylneuraminic acid.
Mentions: Sialic acids are one of the most common terminal monosaccharides found on cell surface glycans of mammals and other higher eukaryotes. Due to its ubiquity and properties sialic acids are involved in many biological functions ranging from early fetal development, cellular recognition and adhesion processes to protein half-life and utilization by influenza virus in entry to cell [1]. The term sialic acid covers more than 50 different glycan structures with a common nine carbon structural backbone and a carboxylic acid group at carbon one. The most common sialic acid structure variant and main metabolic precursor of most other sialic acids is N-Acetylneuraminic acid, Neu5Ac (Figure 1). Cellular Neu5Ac is produced by recycling it from surface glycoproteins using salvage pathway [2] or by de-novo biosynthesis route from other metabolic precursors [3]. The end product of the De novo pathway is activated in the nucleus by N-acylneuraminate cytidylyltransferase to produce CMP-Neu5Ac that can be utilized by the Golgi glycosyltransferases in biosynthesis of glycoconjugates.

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