5-Hydroxytryptamine1A receptor/Gibetagamma stimulates mitogen-activated protein kinase via NAD(P)H oxidase and reactive oxygen species upstream of src in chinese hamster ovary fibroblasts. Biochem J 2000 Apr 01;347 Pt 1(Pt 1):61-7
Date
03/23/2000Pubmed ID
10727402Pubmed Central ID
PMC1220931DOI
10.1042/0264-6021:3470061Scopus ID
2-s2.0-0034177790 (requires institutional sign-in at Scopus site) 67 CitationsAbstract
The hypothesis of this work is that the 'serotonin' or 5-hydroxytryptamine (5-HT)(1A) receptor, which activates the extracellular signal-regulated kinase (ERK) through a G(i)betagamma-mediated pathway, does so through the intermediate actions of reactive oxygen species (ROS). Five criteria were shown to support a key role for ROS in the activation of ERK by the 5-HT(1A) receptor. (1) Antioxidants inhibit activation of ERK by 5-HT. (2) Application of cysteine-reactive oxidant molecules activates ERK. (3) The 5-HT(1A) receptor alters cellular redox properties, and generates both superoxide and hydrogen peroxide. (4) A specific ROS-producing enzyme [NAD(P)H oxidase] is involved in the activation of ERK. (5) There is specificity both in the effects of various chemical oxidizers, and in the putative location of the ROS in the ERK activation pathway. We propose that NAD(P)H oxidase is located in the ERK activation pathway stimulated by the transfected 5-HT(1A) receptor in Chinese hamster ovary (CHO) cells downstream of G(i)betagamma subunits and upstream of or at the level of the non-receptor tyrosine kinase, Src. Moreover, these experiments provide confirmation that the transfected human 5-HT(1A) receptor induces the production of ROS (superoxide and hydrogen peroxide) in CHO cells, and support the possibility that an NAD(P)H oxidase-like enzyme might be involved in the 5-HT-mediated generation of both superoxide and hydrogen peroxide.
Author List
Mukhin YV, Garnovskaya MN, Collinsworth G, Grewal JS, Pendergrass D, Nagai T, Pinckney S, Greene EL, Raymond JRAuthor
John R. Raymond MD President, CEO, Professor in the President department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AcetylcysteineAnimals
CHO Cells
Cricetinae
Fibroblasts
GTP-Binding Protein alpha Subunits, Gi-Go
Humans
Hydrogen Peroxide
Kinetics
Mitogen-Activated Protein Kinases
NADH, NADPH Oxidoreductases
NADPH Oxidases
Oxidation-Reduction
Phosphorylation
Reactive Oxygen Species
Receptors, Serotonin
Receptors, Serotonin, 5-HT1
Recombinant Proteins
Serotonin
Superoxides
Transfection
Virulence Factors, Bordetella
src Homology Domains