Faculty Collaboration Database

The mechanism of transmembrane S-nitrosothiol transport. Proc Natl Acad Sci U S A 2004 May 25;101(21):7891-6

Date

05/19/2004

Pubmed ID

15148403

Pubmed Central ID

PMC419527

DOI

10.1073/pnas.0401167101

Scopus ID

2-s2.0-2542516981 (requires institutional sign-in at Scopus site)   161 Citations

Abstract

S-nitrosothiols have been suggested to play an important role in nitric oxide (NO)-mediated biological events. However, the mechanisms by which an S-nitrosothiol (or the S-nitroso functional group) is transferred across cell membrane are still poorly understood. We have demonstrated previously that the degradation of S-nitrosoglutathione (GSNO) by cells absolutely required the presence of cystine in the extracellular medium and proposed a mechanism that involved the reduction of cystine to cysteine, followed by the reaction of cysteine with GSNO to form S-nitrosocysteine (CysNO), mixed disulfides, and nitrosyl anion. In the present study we have assessed the effect of cystine on the transfer of the S-nitroso functional group from the extracellular to the intracellular space. Using RAW 264.7 cells, we found that the presence of L-cystine enhanced GSNO-dependent S-nitrosothiol uptake, increasing the intracellular S-nitrosothiol level from approximately 60 pmol/mg of protein to approximately 3 nmol/mg of protein. The uptake seems to depend on the reduction of L-cystine to L-cysteine, which involves the xc- amino acid transport system, the transnitrosation between GSNO and L-cysteine to form L-CysNO, and uptake of L-CysNO via amino acid transport system L. Compared with GSNO, (Z)-1-[N-(3-ammoniopropyl)-N-[4-(3-aminopropylammonio)butyl]-amino]diazen-1-ium-1,2-diolate, an NO donor, is much less effective at intracellular S-nitrosothiol formation in the presence of L-cystine or L-cysteine, suggesting that the biochemical changes that occur after exposure of cells to S-nitrosothiol, with respect to thiol chemistry, are distinctly different from those observed with NO.

Author List

Zhang Y, Hogg N

Author

Neil Hogg PhD Associate Dean, Professor in the Biophysics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Biological Transport
Cell Line
Cell Membrane
Cysteine
Cystine
Disulfides
Oxidation-Reduction
S-Nitrosoglutathione
S-Nitrosothiols