EMAIL THIS PAGE TO A FRIEND

Oncology reports

Alteration of O-GlcNAcylation affects serine phosphorylation and regulates gene expression and activity of pyruvate kinase M2 in colorectal cancer cells.


PMID 26252736

Abstract

O-GlcNAcylation is a dynamic post-translational modification that has extensive crosstalk with phosphorylation either at the same or adjacent sites of various proteins. We have previously reported that O-GlcNAcylation level was increased in primary breast and colorectal cancer, but the interplay of the two modifications remains unclear. Therefore, we explored crosstalk of the modifications by RNA interference against O-GlcNAc transferase (OGT) in colorectal cancer cells. Two-dimensional immunoblotting and mass spectrometric analysis showed that the levels of O-GlcNAc and serine phosphorylation of many proteins including serine hydroxymethyltransferase, cytokeratin-8, pyruvate kinase M2 (PKM2), heterogeneous nuclear ribonucleoprotein L, and lamin-B1, were reduced in siOGT cells compared to siScramble cells. In HT29 cells, immunoprecipitated PKM2 revealed decreased O-GlcNAc and serine phosphorylation levels after siOGT knockdown, but increased levels after treatment with Thiamet-G, an inhibitor of O-GlcNAcase (OGA). In addition, when global O-GlcNAcylation was enhanced by treating cells with Thiamet-G, PKM2 expression level was upregulated, but PKM2-specific activity was decreased. On the other hand, in OGT knockdown cells, PKM2 expression level was downregulated, but PKM2-specific activity was increased. Moreover, the metastatic colorectal cancer cells, SW620, had more O-GlcNAc-PKM2 and showed lower PKM2-specific activity compared to the non-metastatic colorectal cancer SW480 cells. These results suggested roles of O-GlcNAcylation in modulating serine phosphorylation, as well as in regulating PKM2 activity and expression. Interfering levels of O-GlcNAcylation of PKM2 may be a novel target in controlling cancer metabolism and tumorigenesis of colorectal cancer.

Related Materials

Product #

Image

Description

Molecular Formula

Add to Cart

SAB2500715
Anti-OGT antibody produced in goat, affinity isolated antibody, buffered aqueous solution
SAB2101676
Anti-OGT antibody produced in rabbit, affinity isolated antibody
HPA030751
Anti-OGT antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution, ab1
HPA030752
Anti-OGT antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution, ab2
HPA029501
Anti-PKM antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
SAB4200105
Anti-PKM2 (C-terminal) antibody produced in rabbit, ~1.5 mg/mL, affinity isolated antibody
SAB4200094
Anti-PKM2 (isoform M1) antibody produced in rabbit, ~1.5 mg/mL, affinity isolated antibody
SAB4200095
Anti-PKM2 (isoform M2) antibody produced in rabbit, ~1.5 mg/mL, affinity isolated antibody
SAB4300662
Anti-PKM2 antibody produced in rabbit, affinity isolated antibody
M8146
Methyl-β-D-thiogalactoside
C7H14O5S
EHU082301 MISSION® esiRNA, esiRNA human OGT (esiRNA1)
EMU006701 MISSION® esiRNA, esiRNA targeting mouse Ogt (esiRNA1)
WH0005315M1
Monoclonal Anti-PKM2 antibody produced in mouse, clone 5D2-3B3, ascites fluid, buffered aqueous solution
SAB1404212
Monoclonal Anti-PKM2, (C-terminal) antibody produced in mouse, clone 2D8, purified immunoglobulin, buffered aqueous solution
P1053
O-Phospho-L-threonine
C4H10NO6P
PLA0286
Rabbit anti-PKM2 Antibody, Affinity Purified, Powered by Bethyl Laboratories, Inc.
730734
Trimethylgallium, packaged for use in deposition systems
C3H9Ga