Differential effects of substrate-analogue inhibitors on nitric oxide synthase dimerization

Kyu Sun Lee, Dong Keon Lee, Dooil Jeoung, Hansoo Lee, Jongseon Choe, Kwon Soo Ha, Moo Ho Won, Young-Guen Kwon, Young Myeong Kim

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Nitric oxide synthase (NOS) isoforms are hemoenzymes that are only active as homodimers. We have examined the effect of the substrate-analogue inhibitors, N G -monomethyl-l-arginine (l-NMA), N G -nitro-l-arginine (l-NNA), N G -nitro-l-arginine methyl ester (l-NAME), N 5 -(1-iminoethyl)-l-ornithine (l-NIO), and N 6 -(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 languageEnglish
Pages (from-to)49-55
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume418
Issue number1
DOIs
Publication statusPublished - 2012 Feb 3

Fingerprint

Dimerization
Nitric Oxide Synthase
Arginine
Substrates
Amidines
Ornithine
Guanidine
Endothelial cells
Dimers
Interferons
Lysine
Lipopolysaccharides
Catalyst activity
Protein Isoforms
Endothelial Cells

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lee, Kyu Sun ; Lee, Dong Keon ; Jeoung, Dooil ; Lee, Hansoo ; Choe, Jongseon ; Ha, Kwon Soo ; Won, Moo Ho ; Kwon, Young-Guen ; Kim, Young Myeong. / Differential effects of substrate-analogue inhibitors on nitric oxide synthase dimerization. In: Biochemical and Biophysical Research Communications. 2012 ; Vol. 418, No. 1. pp. 49-55.
@article{96d6a170e5104c44980c0b648295883f,
title = "Differential effects of substrate-analogue inhibitors on nitric oxide synthase dimerization",
abstract = "Nitric oxide synthase (NOS) isoforms are hemoenzymes that are only active as homodimers. We have examined the effect of the substrate-analogue inhibitors, N G -monomethyl-l-arginine (l-NMA), N G -nitro-l-arginine (l-NNA), N G -nitro-l-arginine methyl ester (l-NAME), N 5 -(1-iminoethyl)-l-ornithine (l-NIO), and N 6 -(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.",
author = "Lee, {Kyu Sun} and Lee, {Dong Keon} and Dooil Jeoung and Hansoo Lee and Jongseon Choe and Ha, {Kwon Soo} and Won, {Moo Ho} and Young-Guen Kwon and Kim, {Young Myeong}",
year = "2012",
month = "2",
day = "3",
doi = "10.1016/j.bbrc.2011.12.123",
language = "English",
volume = "418",
pages = "49--55",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

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-Guen; Kim, Young Myeong.

In: Biochemical and Biophysical Research Communications, Vol. 418, No. 1, 03.02.2012, p. 49-55.

Research output: Contribution to journalArticle

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-Guen

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, N G -monomethyl-l-arginine (l-NMA), N G -nitro-l-arginine (l-NNA), N G -nitro-l-arginine methyl ester (l-NAME), N 5 -(1-iminoethyl)-l-ornithine (l-NIO), and N 6 -(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, N G -monomethyl-l-arginine (l-NMA), N G -nitro-l-arginine (l-NNA), N G -nitro-l-arginine methyl ester (l-NAME), N 5 -(1-iminoethyl)-l-ornithine (l-NIO), and N 6 -(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.

UR - http://www.scopus.com/inward/record.url?scp=84862776579&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84862776579&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2011.12.123

DO - 10.1016/j.bbrc.2011.12.123

M3 - Article

VL - 418

SP - 49

EP - 55

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -