PMID- 15139849
OWN - NLM
STAT- MEDLINE
DCOM- 20041210
LR  - 20181113
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 381
IP  - Pt 3
DP  - 2004 Aug 1
TI  - Oxidative stress inhibits MEKK1 by site-specific glutathionylation in the 
      ATP-binding domain.
PG  - 675-83
AB  - Many intracellular signalling events are accompanied by generation of reactive 
      oxygen species in cells. Oxidation of protein thiol groups is an emerging theme 
      in signal-transduction research. We have found that MEKK1 [MAPK 
      (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) 
      kinase kinase 1], an upstream activator of the SAPK/JNK (stress-activated protein 
      kinase/c-Jun N-terminal kinase) pathway, is directly inhibited by cysteine 
      alkylation using NEM (N-ethylmaleimide). The related kinase, ASK1 (apoptosis 
      signal-regulating kinase 1), was not inhibited, but was instead activated by NEM. 
      Inhibition of MEKK1 requires a single unique cysteine residue (Cys1238) in the 
      ATP-binding domain of MEKK1. Oxidative stress induced by menadione 
      (2-methyl-1,4-naphthoquinone) also inhibited MEKK1, but activated ASK1, in cells. 
      MEKK1 inhibition by menadione also required Cys1238. Oxidant-inhibited MEKK1 was 
      re-activated by dithiothreitol and glutathione, supporting reversible cysteine 
      oxidation as a mechanism. Using various chemical probes, we excluded modification 
      by S-nitrosylation or oxidation of cysteine to sulphenic acid. Oxidant-inhibited 
      MEKK1 migrated normally on non-reducing gels, excluding the possibility of intra- 
      or inter-molecular disulphide bond formation. MEKK1 was inhibited by 
      glutathionylation in vitro, and MEKK1 isolated from menadione-treated cells was 
      shown by MS to be modified by glutathione on Cys1238. Our results support a model 
      whereby the redox environment within the cell selectively regulates stress 
      signalling through MEKK1 versus ASK1, and may thereby participate in the 
      induction of apoptosis by oxidative stress.
FAU - Cross, Janet V
AU  - Cross JV
AD  - Department of Pathology, University of Virginia Medical School, PO Box 800214, 
      Charlottesville, VA 22908-0214, U.S.A.
FAU - Templeton, Dennis J
AU  - Templeton DJ
LA  - eng
GR  - R01 CA066134/CA/NCI NIH HHS/United States
GR  - CA66134/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Oxidants)
RN  - 0 (Peptides)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.11.25 (MAP Kinase Kinase Kinase 1)
RN  - GAN16C9B8O (Glutathione)
RN  - HG18B9YRS7 (Valine)
RN  - K848JZ4886 (Cysteine)
RN  - O3C74ACM9V (Ethylmaleimide)
RN  - T8ID5YZU6Y (Dithiothreitol)
SB  - IM
CIN - Biochem J. 2004 Aug 1;381(Pt 3):e1-2. PMID: 15270699
MH  - Adenosine Triphosphate/*metabolism
MH  - Alkylation
MH  - Amino Acid Sequence/genetics/physiology
MH  - Amino Acid Substitution
MH  - Binding Sites/physiology
MH  - Catalytic Domain/drug effects
MH  - Cell Line, Tumor
MH  - Cysteine/metabolism
MH  - Dithiothreitol/pharmacology
MH  - Enzyme Inhibitors/pharmacology
MH  - Ethylmaleimide/pharmacology
MH  - Glutathione/*metabolism
MH  - Humans
MH  - Lymph Nodes/enzymology/pathology
MH  - MAP Kinase Kinase Kinase 1/*antagonists & 
      inhibitors/chemistry/metabolism/physiology
MH  - Male
MH  - Molecular Sequence Data
MH  - Mutation/physiology
MH  - Oxidants/antagonists & inhibitors/pharmacology
MH  - Oxidation-Reduction
MH  - Oxidative Stress/drug effects/*physiology
MH  - Peptides/chemistry/*metabolism/physiology
MH  - Prostatic Neoplasms/enzymology/pathology
MH  - Protein Structure, Tertiary
MH  - Valine/metabolism
PMC - PMC1133876
EDAT- 2004/05/14 05:00
MHDA- 2004/12/16 09:00
PMCR- 2005/02/01
CRDT- 2004/05/14 05:00
PHST- 2004/05/13 00:00 [accepted]
PHST- 2004/05/11 00:00 [revised]
PHST- 2004/04/09 00:00 [received]
PHST- 2004/05/14 05:00 [pubmed]
PHST- 2004/12/16 09:00 [medline]
PHST- 2004/05/14 05:00 [entrez]
PHST- 2005/02/01 00:00 [pmc-release]
AID - BJ20040591 [pii]
AID - bj3810675 [pii]
AID - 10.1042/BJ20040591 [doi]
PST - ppublish
SO  - Biochem J. 2004 Aug 1;381(Pt 3):675-83. doi: 10.1042/BJ20040591.