Nitric oxide (NO) is a precursor of reactive nitrating species, peroxynitrite and nitrogen dioxide, which modify proteins to generate oxidized species such as 3-nitrotyrosine that has been used as a hallmark of peroxynitrite-mediated oxidative stress on proteins. In the last few years however, a growing body of evidence indicates that NO also regulates a myriad of physiologic responses by modifying tyrosine residues. Looking for the molecular event triggered by NO in nerve growth factor (NGF)-induced neuronal differentiation, we recently reported that in differentiating PC12 cells, the cytoskeleton becomes the main cellular fraction containing nitrotyrosinated proteins, and α-tubulin is the major target. In the present work, we focus on the investigation of the sites of tyrosine nitration in α-tubulin purified by two-dimensional gel electrophoresis following anti-α-tubulin immunoprecipitation of protein extract from NGF-treated PC12 cells. Using Western blotting and matrix-assisted laser desorption/ionization-time of flight analysis, we show for the first time, both in vivo and in vitro, that nitration can occur on α-tubulin at sites other than the C-terminus and we positively identify Tyr 161 and Tyr 357 as two specific amino acids endogenously nitrated.

Nitroproteome of neuron-like PC12 cells / G. Tedeschi - In: Sezione Italiana Human Proteome Organization (IHUPO) : 3. Italian Human Proteome Organization National Meeting : 29 September-1 October 2005, Lodi, Italy / Italian Human Proteome Organization. - [S.n.] : Italian Human Proteome Organization, 2005. - pp. 42-42 (( Intervento presentato al 3. convegno Proteomics : from new methodologies to cellular mechanisms tenutosi a Lodi nel 2005.

Nitroproteome of neuron-like PC12 cells

G. Tedeschi
Primo
2005

Abstract

Nitric oxide (NO) is a precursor of reactive nitrating species, peroxynitrite and nitrogen dioxide, which modify proteins to generate oxidized species such as 3-nitrotyrosine that has been used as a hallmark of peroxynitrite-mediated oxidative stress on proteins. In the last few years however, a growing body of evidence indicates that NO also regulates a myriad of physiologic responses by modifying tyrosine residues. Looking for the molecular event triggered by NO in nerve growth factor (NGF)-induced neuronal differentiation, we recently reported that in differentiating PC12 cells, the cytoskeleton becomes the main cellular fraction containing nitrotyrosinated proteins, and α-tubulin is the major target. In the present work, we focus on the investigation of the sites of tyrosine nitration in α-tubulin purified by two-dimensional gel electrophoresis following anti-α-tubulin immunoprecipitation of protein extract from NGF-treated PC12 cells. Using Western blotting and matrix-assisted laser desorption/ionization-time of flight analysis, we show for the first time, both in vivo and in vitro, that nitration can occur on α-tubulin at sites other than the C-terminus and we positively identify Tyr 161 and Tyr 357 as two specific amino acids endogenously nitrated.
α-Tubulin; Cytoskeleton; Matrix-assisted laser desorption/ionization-time of flight; Neural cells; Nitric oxide
Settore BIO/10 - Biochimica
Book Part (author)
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/15495
Citazioni
  • ???jsp.display-item.citation.pmc??? 12
  • Scopus 45
  • ???jsp.display-item.citation.isi??? 42
social impact