title: MARCKS protein mediates hydrogen peroxide regulation of endothelial permeability.
abstract: Impairment of endothelial barrier function is implicated in many vascular and inflammatory disorders. One prevalent mechanism of endothelial dysfunction is an increase in reactive oxygen species under oxidative stress. Previous reports have demonstrated that hydrogen peroxide (H(2)O(2)), a highly stable reactive oxygen species that modulates physiological signaling pathways, also enhances endothelial permeability, but the mechanism of this effect is unknown. Here, we identify the actin-binding protein myristoylated alanine-rich C-kinase substrate (MARCKS) as a key mediator of the H(2)O(2)-induced permeability change in bovine aortic endothelial cells. MARCKS knockdown and H(2)O(2) treatment alter the architecture of the actin cytoskeleton in endothelial cells, and H(2)O(2) induces the phosphorylation and translocation of MARCKS from the cell membrane to the cytosol. Using pharmacological inhibitors and small interference RNA constructs directed against specific proteins, we uncover a signaling cascade from Rac1 to Abl1, phospholipase Cγ1, and PKCδ that is triggered by H(2)O(2) and leads to MARCKS phosphorylation. Our findings establish a distinct role for MARCKS in the regulation of H(2)O(2)-induced permeability change in endothelial cells, and suggest potential new therapeutic targets for the treatment of disorders involving oxidative stress and altered endothelial permeability.
authors.affiliation: Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Benjamin Y
authors.initials: BY
authors.lastname: Jin
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Lin
authors.forename: David E
authors.initials: DE
authors.lastname: Golan
authors.forename: Thomas
authors.initials: T
authors.lastname: Michel
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000166:cytology; D002199:Capillary Permeability / Q000502:physiology*; D002417:Cattle; D042783:Endothelial Cells / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006861:Hydrogen Peroxide / Q000378:metabolism*; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D018384:Oxidative Stress / Q000502:physiology*; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D006861:Hydrogen Peroxide