Phosphoglycerate Mutase-derived Polypeptide Inhibits Glycolytic Flux and Induces Cell Growth Arrest in Tumor Cell Lines

Abstract

The putative tumor metastasis suppressor protein Nm23-H1 is a nucleoside diphosphate kinase that exhibits a novel protein kinase activity when bound to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In this study we show that the glycolytic enzyme phosphoglycerate mutase B (PGM) becomes phosphorylated in the presence of the Nm23-H1⅐GAPDH complex in vitro. Mutation of His-10 in PGM abolishes the Nm23-H1⅐GAPDH complex-induced phosphorylation. Nm23-H1, GAPDH, and PGM are known to co-localize as shown by free flow isoelectric focusing. In association with Nm23-H1 and GAPDH, PGM could be activated by dCTP, which is a substrate of Nm23-H1, in addition to the well known PGM activator 2,3-bisphosphoglycerate. A synthetic cell-penetrating peptide (PGMtide) encompassing the phosphorylated histidine and several residues from PGM (LIRHGE) promoted growth arrest of several tumor cell lines, whereas proliferation of tested non-tumor cells was not influenced. Analysis of metabolic activity of one of the tumor cell lines, MCF-7, indicated that PGMtide inhibited glycolytic flux, consistent with in vivo inhibition of PGM. The specificity of the observed effect was further determined experimentally by testing the effect of PGMtide on cells growing in the presence of pyruvate, which helps to compensate PGM inhibition in the glycolytic pathway. Thus, growth of MCF-7 cells was not arrested by PGMtide in the presence of pyruvate. The data presented here provide evidence that inhibition of PGM activity can be achieved by exogenous addition of a polypeptide, resulting in inhibition of glycolysis and cell growth arrest in cell culture.

CRHR1-dependent effects on protein expression and posttranslational modification in AtT-20 cells

“…It has been reported that PGAM1 can also be activated by dCTP in the presence of NME1, a nucleoside diphosphate kinase (Engel et al, 2004). In our analysis we detected a regulation of NME1 at the protein expression level before PGAM1 appears to be modified.…”

Regulation of Glycolytic Enzyme Phosphoglycerate Mutase-1 by Sirt1 Protein-mediated Deacetylation

“…Either 3KQ, 3KR, or wild type PGAM was overexpressed in HEK293 cells cultured in pyruvate-free media, and the concentrations of glucose, lactate, and alanine were measured. Growth in pyruvate-free media simplifies measurements of glycolytic flux (34). Compared with cells expressing PGAM and the 3KR variant, cells expressing 3KQ exhibited stimulated glucose consumption and a corresponding increase in lactate levels (Fig.…”

Comparative Transcriptomic and Proteomic Analyses of Trichomonas vaginalis following Adherence to Fibronectin

“…Important for glycolytic flux in cells is phosphoglycerate mutase (21), which may play a similar role within trichomonads. Finally, there are numerous carbohydratedegrading and/or -modifying enzymes, some of which are highly expressed in the FN-EST transcriptome, and these enzymes may play an important role in aborting any lectin-type interactions between the parasite and the host cell.…”