Phosphodiesterases (PDEs) are a superfamily of evolutionarily conserved cyclic nucleotide (cAMP/cGMP)-hydrolyzing enzymes, components of transduction pathways regulating crucial aspects of cell life. Within this family, the cGMP-dependent PDE5 is the major hydrolyzing enzyme in many mammalian tissues, where it regulates a number of cellular and tissular processes. Using Kluyveromyces lactis as a model organism, the murine PDE5A1, A2 and A3 isoforms were successfully expressed and studied, evidencing, for the first time, a distinct role of each isoform in the control, modulation and maintenance of the cellular redox metabolism. Moreover, we demonstrated that the short N-terminal peptide is responsible for the tetrameric assembly of MmPDE5A1 and for the mito- chondrial localization of MmPDE5A2. We also analyzed MmPDE5A1, A2 and A3 using small-angle X-ray scattering (SAXS), transmission electron microscopy (TEM), structural mass spectrometry (MS) and polyacrylamide gel electrophoresis in their native conditions (native-PAGE) and in the presence of redox agents. These analyses pointed towards the role of a few specific cysteines in the isoforms’ oligomeric assembly and the loss of enzymatic activity when modified.

Structural characterization of murine phosphodiesterase 5 isoforms and involvement of cysteine residues in supramolecular assembly / Giorgi, Mauro; Miele, Adriana Erica; Cardarelli, Silvia; Giorgi, Alessandra; Massimi, Mara; Biagioni, Stefano; Saliola, Michele. - In: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES. - ISSN 1422-0067. - 24:2(2023). [10.3390/ijms24021108]

Structural characterization of murine phosphodiesterase 5 isoforms and involvement of cysteine residues in supramolecular assembly

Mauro Giorgi
;
Adriana Erica Miele
;
Silvia Cardarelli;Alessandra Giorgi;Stefano Biagioni;Michele Saliola
2023

Abstract

Phosphodiesterases (PDEs) are a superfamily of evolutionarily conserved cyclic nucleotide (cAMP/cGMP)-hydrolyzing enzymes, components of transduction pathways regulating crucial aspects of cell life. Within this family, the cGMP-dependent PDE5 is the major hydrolyzing enzyme in many mammalian tissues, where it regulates a number of cellular and tissular processes. Using Kluyveromyces lactis as a model organism, the murine PDE5A1, A2 and A3 isoforms were successfully expressed and studied, evidencing, for the first time, a distinct role of each isoform in the control, modulation and maintenance of the cellular redox metabolism. Moreover, we demonstrated that the short N-terminal peptide is responsible for the tetrameric assembly of MmPDE5A1 and for the mito- chondrial localization of MmPDE5A2. We also analyzed MmPDE5A1, A2 and A3 using small-angle X-ray scattering (SAXS), transmission electron microscopy (TEM), structural mass spectrometry (MS) and polyacrylamide gel electrophoresis in their native conditions (native-PAGE) and in the presence of redox agents. These analyses pointed towards the role of a few specific cysteines in the isoforms’ oligomeric assembly and the loss of enzymatic activity when modified.
2023
PDE5; SAXS; TEM; MS; native-PAGE; quaternary structure; reactive Cys
01 Pubblicazione su rivista::01a Articolo in rivista
Structural characterization of murine phosphodiesterase 5 isoforms and involvement of cysteine residues in supramolecular assembly / Giorgi, Mauro; Miele, Adriana Erica; Cardarelli, Silvia; Giorgi, Alessandra; Massimi, Mara; Biagioni, Stefano; Saliola, Michele. - In: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES. - ISSN 1422-0067. - 24:2(2023). [10.3390/ijms24021108]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1677555
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