Abstract
Nitric oxide synthase (NOS) isoforms are hemoenzymes that are only active as homodimers. We have examined the effect of the substrate-analogue inhibitors, NG-monomethyl-l-arginine (l-NMA), NG-nitro-l-arginine (l-NNA), NG-nitro-l-arginine methyl ester (l-NAME), N5-(1-iminoethyl)-l-ornithine (l-NIO), and N6-(1-iminoethyl)-l-lysine (l-NIL), the guanidine-containing inhibitor aminoguanidine (AG), and the amidine moiety-containing iNOS-specific inhibitor 1400W, on the formation of NOS dimer. Of these inhibitors, l-NMA effectively not only inhibited iNOS dimerization, but also destabilized its dimeric form in RAW264.7 cells stimulated with lipopolysaccharide plus interferon-γ, but not eNOS dimerization in endothelial cells. Importantly, this inhibition was highly correlated with NO production. These inhibitory effects were significantly reversed by addition of l-arginine. However, l-NNA, l-NAME, and AG in part or significantly increased dimerization of iNOS and eNOS in intact cells, and the other inhibitors assessed did not alter dimerization of iNOS and eNOS. These data taken together suggest that substituted groups of an arginine guanidino moiety play an important role in NOS dimerization as well as its catalytic activity. Our results indicate that l-NMA can inhibit iNOS-dependent NO production by preventing iNOS dimerization and destabilizing its dimeric form.
| Original language | English |
|---|---|
| Pages (from-to) | 49-55 |
| Number of pages | 7 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 418 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2012 Feb 3 |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
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Differential effects of substrate-analogue inhibitors on nitric oxide synthase dimerization. / Lee, Kyu Sun; Lee, Dong Keon; Jeoung, Dooil; Lee, Hansoo; Choe, Jongseon; Ha, Kwon Soo; Won, Moo Ho; Kwon, Young Geun; Kim, Young Myeong.
In: Biochemical and Biophysical Research Communications, Vol. 418, No. 1, 03.02.2012, p. 49-55.Research output: Contribution to journal › Article
TY - JOUR
T1 - Differential effects of substrate-analogue inhibitors on nitric oxide synthase dimerization
AU - Lee, Kyu Sun
AU - Lee, Dong Keon
AU - Jeoung, Dooil
AU - Lee, Hansoo
AU - Choe, Jongseon
AU - Ha, Kwon Soo
AU - Won, Moo Ho
AU - Kwon, Young Geun
AU - Kim, Young Myeong
PY - 2012/2/3
Y1 - 2012/2/3
N2 - Nitric oxide synthase (NOS) isoforms are hemoenzymes that are only active as homodimers. We have examined the effect of the substrate-analogue inhibitors, NG-monomethyl-l-arginine (l-NMA), NG-nitro-l-arginine (l-NNA), NG-nitro-l-arginine methyl ester (l-NAME), N5-(1-iminoethyl)-l-ornithine (l-NIO), and N6-(1-iminoethyl)-l-lysine (l-NIL), the guanidine-containing inhibitor aminoguanidine (AG), and the amidine moiety-containing iNOS-specific inhibitor 1400W, on the formation of NOS dimer. Of these inhibitors, l-NMA effectively not only inhibited iNOS dimerization, but also destabilized its dimeric form in RAW264.7 cells stimulated with lipopolysaccharide plus interferon-γ, but not eNOS dimerization in endothelial cells. Importantly, this inhibition was highly correlated with NO production. These inhibitory effects were significantly reversed by addition of l-arginine. However, l-NNA, l-NAME, and AG in part or significantly increased dimerization of iNOS and eNOS in intact cells, and the other inhibitors assessed did not alter dimerization of iNOS and eNOS. These data taken together suggest that substituted groups of an arginine guanidino moiety play an important role in NOS dimerization as well as its catalytic activity. Our results indicate that l-NMA can inhibit iNOS-dependent NO production by preventing iNOS dimerization and destabilizing its dimeric form.
AB - Nitric oxide synthase (NOS) isoforms are hemoenzymes that are only active as homodimers. We have examined the effect of the substrate-analogue inhibitors, NG-monomethyl-l-arginine (l-NMA), NG-nitro-l-arginine (l-NNA), NG-nitro-l-arginine methyl ester (l-NAME), N5-(1-iminoethyl)-l-ornithine (l-NIO), and N6-(1-iminoethyl)-l-lysine (l-NIL), the guanidine-containing inhibitor aminoguanidine (AG), and the amidine moiety-containing iNOS-specific inhibitor 1400W, on the formation of NOS dimer. Of these inhibitors, l-NMA effectively not only inhibited iNOS dimerization, but also destabilized its dimeric form in RAW264.7 cells stimulated with lipopolysaccharide plus interferon-γ, but not eNOS dimerization in endothelial cells. Importantly, this inhibition was highly correlated with NO production. These inhibitory effects were significantly reversed by addition of l-arginine. However, l-NNA, l-NAME, and AG in part or significantly increased dimerization of iNOS and eNOS in intact cells, and the other inhibitors assessed did not alter dimerization of iNOS and eNOS. These data taken together suggest that substituted groups of an arginine guanidino moiety play an important role in NOS dimerization as well as its catalytic activity. Our results indicate that l-NMA can inhibit iNOS-dependent NO production by preventing iNOS dimerization and destabilizing its dimeric form.
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U2 - 10.1016/j.bbrc.2011.12.123
DO - 10.1016/j.bbrc.2011.12.123
M3 - Article
C2 - 22240022
AN - SCOPUS:84862776579
VL - 418
SP - 49
EP - 55
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -