Phosphoglycerate mutase-derived polypeptide inhibits glycolytic flux and induces cell growth arrest in tumor cell lines.
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-H1.GAPDH complex in vitro. Mutation of His-10 in PGM abolishes the Nm23-H1.GAPDH 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.
Engel
Matthias
M
Department of Human Genetics, Medical School, University of Saarland, Building 60, D-66421 Homburg, Germany. m.engel@phosphosites.com
Mazurek
Sybille
S
Eigenbrodt
Erich
E
Welter
Cornelius
C
D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000378:metabolism*; D006019:Glycolysis / Q000187:drug effects; D006131:Growth Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D010736:Phosphoglycerate Mutase / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D010766:Phosphorylation; D011485:Protein Binding; D025521:Tumor Suppressor Proteins / Q000737:chemistry* / Q000378:metabolism
D006131:Growth Inhibitors; D010455:Peptides; D025521:Tumor Suppressor Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D009701:Nucleoside-Diphosphate Kinase; D010736:Phosphoglycerate Mutase