title: A novel anti-inflammatory peptide inhibits endothelial cell cytoskeletal rearrangement, nitric oxide synthase translocation, and paracellular permeability increases.
abstract: The endothelial cell (EC) membrane-cytoskeletal interface in part maintains plasma membrane integrity and promotes cell-cell apposition. Nonmuscle filamin (ABP-280), an actin crosslinking protein, promotes orthogonal branching of F-actin and is the major protein that links the peripheral actin network to the plasma membrane through its C-terminal glycoprotein binding site. In response to bradykinin, filamin translocates from the cell periphery to the cytosol within 1 min. A synthetic peptide, corresponding to filamin's C-terminal calcium/calmodulin-dependent protein kinase II phosphorylation site (CaM peptide), prevents calcium-activated filamin translocation in permeabilized bovine pulmonary artery EC. The myristoylated permeable form of this peptide inhibits bradykinin-induced filamin translocation and F-actin rearrangement in cultured intact ECs. In addition, bradykinin-induced paracellular gap formation is significantly attenuated by CaM peptide, which suggests that the presence of a filamin-based peripheral F-actin network is essential for maintaining EC barrier function. Moreover, CaM peptide reduces wound-induced EC migration rate by 40%, which indicates that F-actin rearrangement is required for efficient cell motility. The CaM peptide affects other bradykinin-induced inflammatory responses. EC nitric oxide synthase (eNOS) translocates from the cell membrane to the nuclear fraction within 1-2 min of bradykinin treatment. Pretreatment with CaM peptide inhibits eNOS translocation. However, the peptide has no effect on bradykinin-induced von Willebrand Factor release. In summary, the CaM peptide exhibits several anti-inflammatory properties that include maintaining EC junctional stability and inhibiting eNOS translocation.
authors.affiliation: Microvascular Research Laboratory, Boston University, Massachusetts 02215, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Wang
authors.forename: W F
authors.initials: WF
authors.lastname: Patton
authors.forename: H B
authors.initials: HB
authors.lastname: Hechtman
authors.forename: D
authors.initials: D
authors.lastname: Shepro
mesh_terms: D000199:Actins / Q000187:drug effects / Q000502:physiology; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D001920:Bradykinin / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000494:pharmacology; D002199:Capillary Permeability / Q000187:drug effects*; D002417:Cattle; D002465:Cell Movement / Q000187:drug effects; D003285:Contractile Proteins / Q000494:pharmacology*; D003599:Cytoskeleton / Q000648:ultrastructure*; D004730:Endothelium, Vascular / Q000648:ultrastructure*; D004790:Enzyme Induction; D064448:Filamins; D017629:Gap Junctions / Q000187:drug effects / Q000502:physiology; D008840:Microfilament Proteins / Q000494:pharmacology*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology*; D014945:Wound Healing / Q000187:drug effects; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D000199:Actins; D000893:Anti-Inflammatory Agents; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D014841:von Willebrand Factor; D019001:Nitric Oxide Synthase; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D001920:Bradykinin