Role of ADP ribosylation factor6- Cytohesin1-PhospholipaseD signaling axis in U46619 induced activation of NADPH oxidase in pulmonary artery smooth muscle cell membrane.

• 影响指数: 4.114
• 发表时间: 11 2017 1
• 来源期刊: Arch Biochem Biophys
• DOI: 10.1016/j.abb.2017.08.012
• PMID: 28822840
• 文章类型: Journal Article
• 中科院分区: Q2
• 方向: 生物
• 作者列表: Chakraborti S,Sarkar J,Chowdhury A,Chakraborti T

关键词: ADP rybosylation factor; Cytohesin; NADPH oxidase; Phospholipase D; Pulmonary artery smooth muscle cells; Thromboxane A(2)

Mesh: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / pharmacology; ADP Ribose Transferases / pharmacology; ADP-Ribosylation Factor 6; ADP-Ribosylation Factors / genetics metabolism; Acetophenones / pharmacology; Antioxidants / pharmacology; Botulinum Toxins / pharmacology; Brefeldin A / pharmacology; Bridged Bicyclo Compounds, Heterocyclic; Cell Membrane / drug effects metabolism; Fatty Acids, Unsaturated; GTPase-Activating Proteins / antagonists & inhibitors genetics metabolism; Gene Expression Regulation; Guanine Nucleotide Exchange Factors / antagonists & inhibitors genetics metabolism; Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology; Humans; Hydrazines / pharmacology; Myocytes, Smooth Muscle / cytology drug effects metabolism; NADPH Oxidases / genetics metabolism; Phospholipase D / antagonists & inhibitors genetics metabolism; Primary Cell Culture; Protein Synthesis Inhibitors / pharmacology; Pulmonary Artery / cytology drug effects metabolism; Receptors, Thromboxane A2, Prostaglandin H2 / antagonists & inhibitors genetics metabolism; Signal Transduction; Triazoles / pharmacology; Vasoconstrictor Agents / pharmacology

Abstract

Treatment of human pulmonary artery smooth muscle cells (HPASMCs) with the thromboxane A2 receptor antagonist, SQ29548 inhibited U46619 stimulation of phospholipase D (PLD) and NADPH oxidase activities in the cell membrane. Pretreatment with apocynin inhibited U46619 induced increase in NADPH oxidase activity. The cell membrane contains predominantly PLD2 along with PLD1 isoforms of PLD. Pretreatment with pharmacological and genetic inhibitors of PLD2, but not PLD1, attenuated U46619 stimulation of NADPH oxidase activity. U46619 stimulation of PLD and NADPH oxidase activities were insensitive to BFA and Clostridium botulinum C3 toxin; however, pretreatment with secinH3 inhibited U46619 induced increase in PLD and NADPH oxidase activities suggesting a major role of cytohesin in U46619-induced increase in PLD and NADPH oxidase activities. Arf-1, Arf-6, cytohesin-1 and cytohesin-2 were observed in the cytosolic fraction, but only Arf-6 and cytohesin-1 were translocated to the cell membrane upon treatment with U46619. Coimmunoprecipitation study showed association of Arf-6 with cytohesin-1 in the cell membrane fraction. In vitro binding of GTPγS with Arf-6 required the presence of cytohesin-1 and that occurs in BFA insensitive manner. Overall, BFA insensitive Arf6-cytohesin1 signaling axis plays a pivotal role in U46619-mediated activation of PLD leading to stimulation of NADPH oxidase activity in HPASMCs.