Inhibition of Azotobacter vinelandii rhodanese by NO-donors

Andrea Spallarossa, Fabio Forlani, Silvia Pagani, Luca Salvati, Paolo Visca, Paolo Ascenzi, Martino Bolognesi, Domenico Bordo

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Nitric oxide (NO) is a versatile regulatory molecule that affects enzymatic activity through chemical modification of reactive amino acid residues (e.g., Cys and Tyr) and by binding to metal centers. In the present study, the inhibitory effect of the NO-donors S-nitroso-glutathione (GSNO), (±)E-4-ethyl-2-[E-hydroxyimino]-5-nitro-3-hexenamide (NOR-3), and S-nitroso-N-acetyl-penicillamine (SNAP) on the catalytic activity of Azotobacter vinelandii rhodanese (RhdA) has been investigated. GSNO, NOR-3, and SNAP inhibit RhdA sulfurtransferase activity in a concentration- and time-dependent fashion. The absorption spectrum of the NOR-3-treated RhdA displays a maximum at 335 nm, indicating NO-mediated S-nitrosylation. RhdA inhibition by NO-donors correlates with S-nitrosothiol formation. The reducing agent dithiothreitol prevents RhdA inhibition by NO-donors, fully restores the catalytic activity, and reverts the NOR-3-induced RhdA absorption spectrum to that of the active enzyme. These results indicate that RhdA inhibition occurs via NO-mediated S-nitrosylation of the unique Cys230 catalytic residue.

Original languageEnglish
Pages (from-to)1002-1007
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume306
Issue number4
DOIs
Publication statusPublished - Jul 11 2003

Fingerprint

Thiosulfate Sulfurtransferase
Azotobacter vinelandii
Nitric Oxide Donors
Nitric Oxide
Penicillamine
Absorption spectra
Catalyst activity
S-Nitrosothiols
Dithiothreitol
Reducing Agents
Chemical modification
Glutathione
Metals
Amino Acids
Molecules
Enzymes

Keywords

  • Enzyme inhibition
  • NO-donors
  • Phosphatase
  • Rhodanese
  • Sulfurtransferase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Inhibition of Azotobacter vinelandii rhodanese by NO-donors. / Spallarossa, Andrea; Forlani, Fabio; Pagani, Silvia; Salvati, Luca; Visca, Paolo; Ascenzi, Paolo; Bolognesi, Martino; Bordo, Domenico.

In: Biochemical and Biophysical Research Communications, Vol. 306, No. 4, 11.07.2003, p. 1002-1007.

Research output: Contribution to journalArticle

Spallarossa, A, Forlani, F, Pagani, S, Salvati, L, Visca, P, Ascenzi, P, Bolognesi, M & Bordo, D 2003, 'Inhibition of Azotobacter vinelandii rhodanese by NO-donors', Biochemical and Biophysical Research Communications, vol. 306, no. 4, pp. 1002-1007. https://doi.org/10.1016/S0006-291X(03)01067-2
Spallarossa A, Forlani F, Pagani S, Salvati L, Visca P, Ascenzi P et al. Inhibition of Azotobacter vinelandii rhodanese by NO-donors. Biochemical and Biophysical Research Communications. 2003 Jul 11;306(4):1002-1007. https://doi.org/10.1016/S0006-291X(03)01067-2
Spallarossa, Andrea ; Forlani, Fabio ; Pagani, Silvia ; Salvati, Luca ; Visca, Paolo ; Ascenzi, Paolo ; Bolognesi, Martino ; Bordo, Domenico. / Inhibition of Azotobacter vinelandii rhodanese by NO-donors. In: Biochemical and Biophysical Research Communications. 2003 ; Vol. 306, No. 4. pp. 1002-1007.
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