The phosphoglycerate kinase 1 variants found in carcinoma cells display different catalytic activity and conformational stability compared to the native enzyme

Annarita Fiorillo; Maria Petrosino; Andrea Ilari; Alessandra Pasquo; Alessandra Cipollone; Maristella Maggi; Roberta Chiaraluce; Valerio Consalvi

doi:10.1371/journal.pone.0199191

The phosphoglycerate kinase 1 variants found in carcinoma cells display different catalytic activity and conformational stability compared to the native enzyme

PLoS One. 2018 Jul 11;13(7):e0199191. doi: 10.1371/journal.pone.0199191. eCollection 2018.

Authors

Annarita Fiorillo¹, Maria Petrosino¹, Andrea Ilari², Alessandra Pasquo³, Alessandra Cipollone¹, Maristella Maggi⁴, Roberta Chiaraluce¹, Valerio Consalvi¹

Affiliations

¹ Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome, Italy.
² CNR-Institute of Molecular Biology and Pathology, Rome, Italy.
³ ENEA CR Frascati, Diagnostics and Metrology Laboratory, FSN-TECFIS-DIM, Frascati, Italy.
⁴ Department of Molecular Medicine, Unit of Immunology and General Pathology, University of Pavia, Pavia, Italy.

Abstract

Cancer cells are able to survive in difficult conditions, reprogramming their metabolism according to their requirements. Under hypoxic conditions they shift from oxidative phosphorylation to aerobic glycolysis, a behavior known as Warburg effect. In the last years, glycolytic enzymes have been identified as potential targets for alternative anticancer therapies. Recently, phosphoglycerate kinase 1 (PGK1), an ubiquitous enzyme expressed in all somatic cells that catalyzes the seventh step of glycolysis which consists of the reversible phosphotransfer reaction from 1,3-bisphosphoglycerate to ADP, has been discovered to be overexpressed in many cancer types. Moreover, several somatic variants of PGK1 have been identified in tumors. In this study we analyzed the effect of the single nucleotide variants found in cancer tissues on the PGK1 structure and function. Our results clearly show that the variants display a decreased catalytic efficiency and/or thermodynamic stability and an altered local tertiary structure, as shown by the solved X-ray structures. The changes in the catalytic properties and in the stability of the PGK1 variants, mainly due to the local changes evidenced by the X-ray structures, suggest also changes in the functional role of PGK to support the biosynthetic need of the growing and proliferating tumour cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Diphosphate / chemistry*
Adenosine Diphosphate / metabolism
Amino Acid Sequence
Binding Sites
Cloning, Molecular
Crystallography, X-Ray
Enzyme Stability
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Genetic Vectors / chemistry
Genetic Vectors / metabolism
Glyceric Acids / chemistry*
Glyceric Acids / metabolism
Humans
Kinetics
Models, Molecular
Mutation
Neoplasm Proteins / chemistry*
Neoplasm Proteins / genetics
Neoplasm Proteins / metabolism
Phosphoglycerate Kinase / chemistry*
Phosphoglycerate Kinase / genetics
Phosphoglycerate Kinase / metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Protein Structure, Tertiary
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Substrate Specificity
Thermodynamics

Substances

Glyceric Acids
Neoplasm Proteins
Recombinant Proteins
Adenosine Diphosphate
3-phosphoglycerate
PGK1 protein, human
Phosphoglycerate Kinase

Grants and funding

This study was supported by Prot. FILAS-RU-2014-1020 Regione Lazio, Grant recipient: Valerio Consalvi. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.