title: S-nitrosation of cellular proteins by NO donors in rat embryonic fibroblast 3Y1 cells: factors affecting S-nitrosation.
abstract: The mechanism of protein S-nitrosation in cells is not fully understood. Using rat 3Y1 cells, we addressed this issue. Among S-nitrosothiols and NO donors tested, only S-nitrosocysteine (CysNO) induced S-nitrosation when exposed in Hanks' balanced salt solution (HBSS) and not in serum-containing general culture medium. In HBSS, NO release from CysNO was almost completely abolished by sequestering metal ions with a metal chelator without affecting cellular S-nitrosation. In contrast, L-leucine, a substrate of L-type amino acid transporters (LATs), significantly inhibited S-nitrosation. The absence of S-nitrosation with CysNO in general culture medium resulted not only from a competition with amino acids in the medium for LATs but also from transnitrosation of cysteine residues in serum albumin. Collectively, these results suggest that in simple buffered saline, CysNO-dependent S-nitrosation occurs through a cellular incorporation-dependent mechanism, but if it occurs in general culture media, it may be through an NO-dependent mechanism.
authors: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, Japan.
authors: Norihiro
authors: N
authors: Ryuman
authors: Nobuo
authors: N
authors: Watanabe
authors: Takao
authors: T
authors: Arai
mesh_terms: D026905:Amino Acid Transport Systems / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D003545:Cysteine / Q000031:analogs & derivatives / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000378:metabolism*; D015538:Nitrosation / Q000502:physiology*; D051381:Rats; D026403:S-Nitrosothiols / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism
chemical_list: D026905:Amino Acid Transport Systems; D020030:Nitric Oxide Donors; D026403:S-Nitrosothiols; D012709:Serum Albumin; C021315:S-nitrosocysteine; D007930:Leucine; D003545:Cysteine