Human serine racemase is inhibited by glyceraldehyde 3-phosphate, but not by glyceraldehyde 3-phosphate dehydrogenase

Annalisa Michielon; Francesco Marchesani; Serena Faggiano; Roberta Giaccari; Barbara Campanini; Stefano Bettati; Andrea Mozzarelli; Stefano Bruno

doi:10.1016/j.bbapap.2020.140544

Human serine racemase is inhibited by glyceraldehyde 3-phosphate, but not by glyceraldehyde 3-phosphate dehydrogenase

Biochim Biophys Acta Proteins Proteom. 2021 Jan;1869(1):140544. doi: 10.1016/j.bbapap.2020.140544. Epub 2020 Sep 22.

Authors

Annalisa Michielon¹, Francesco Marchesani¹, Serena Faggiano², Roberta Giaccari¹, Barbara Campanini¹, Stefano Bettati³, Andrea Mozzarelli², Stefano Bruno⁴

Affiliations

¹ Dipartimento di Scienze degli Alimenti e del Farmaco, Università degli Studi di Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy.
² Dipartimento di Scienze degli Alimenti e del Farmaco, Università degli Studi di Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy; Istituto di Biofisica, CNR, Via G. Moruzzi 1, 56124 Pisa, Italy.
³ Dipartimento di Medicina e Chirurgia, Via Volturno 39, 43125 Parma, Italy.
⁴ Dipartimento di Scienze degli Alimenti e del Farmaco, Università degli Studi di Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy. Electronic address: stefano.bruno@unipr.it.

PMID: 32971286
DOI: 10.1016/j.bbapap.2020.140544

Abstract

Murine serine racemase (SR), the enzyme responsible for the biosynthesis of the neuromodulator d-serine, was reported to form a complex with glyceraldehyde 3-phosphate dehydrogenase (GAPDH), resulting in SR inhibition. In this work, we investigated the interaction between the two human orthologues. We were not able to observe neither the inhibition nor the formation of the SR-GAPDH complex. Rather, hSR is inhibited by the hGAPDH substrate glyceraldehyde 3-phosphate (G3P) in a time- and concentration-dependent fashion, likely through a covalent reaction of the aldehyde functional group. The inhibition was similar for the two G3P enantiomers but it was not observed for structurally similar aldehydes. We ruled out a mechanism of inhibition based on the competition with either pyridoxal phosphate (PLP) - described for other PLP-dependent enzymes when incubated with small aldehydes - or ATP. Nevertheless, the inhibition time course was affected by the presence of hSR allosteric and orthosteric ligands, suggesting a conformation-dependence of the reaction.

Keywords: Enzyme inhibition; Glyceraldehyde 3-phosphate dehydrogenase; Protein-protein interaction; Serine racemase.

MeSH terms

2,3-Diphosphoglycerate / chemistry
2,3-Diphosphoglycerate / metabolism
Adenosine Triphosphate / chemistry*
Adenosine Triphosphate / metabolism
Aldehydes / chemistry
Aldehydes / metabolism
Catalytic Domain
Cloning, Molecular
Enzyme Inhibitors / chemistry*
Enzyme Inhibitors / metabolism
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Genetic Vectors / chemistry
Genetic Vectors / metabolism
Glyceraldehyde / chemistry
Glyceraldehyde / metabolism
Glyceraldehyde 3-Phosphate / chemistry*
Glyceraldehyde 3-Phosphate / metabolism
Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry*
Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
Humans
Kinetics
Models, Molecular
Protein Binding
Pyridoxal Phosphate / chemistry*
Pyridoxal Phosphate / metabolism
Racemases and Epimerases / antagonists & inhibitors
Racemases and Epimerases / chemistry*
Racemases and Epimerases / genetics
Racemases and Epimerases / metabolism
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Stereoisomerism
Substrate Specificity

Substances

Aldehydes
Enzyme Inhibitors
Recombinant Proteins
2,3-Diphosphoglycerate
Glyceraldehyde 3-Phosphate
Glyceraldehyde
Pyridoxal Phosphate
Adenosine Triphosphate
propionaldehyde
Glyceraldehyde-3-Phosphate Dehydrogenases
Racemases and Epimerases
serine racemase