title: Binding of ADAM28 to P-selectin glycoprotein ligand-1 enhances P-selectin-mediated leukocyte adhesion to endothelial cells.
abstract: ADAMs (a disintegrin and metalloproteinases) are a recently discovered gene family of multifunctional proteins with the disintegrin-like and metalloproteinase domains. To analyze the biological functions of ADAM28, we screened binding molecules to secreted-type ADAM28 (ADAM28s) by the yeast two-hybrid system and identified P-selectin glycoprotein ligand-1 (PSGL-1). Binding between the disintegrin-like domain of ADAM28s and the extracellular portion of PSGL-1 was determined by yeast two-hybrid assays, binding assays of the domain-specific recombinant ADAM28s species using PSGL-1 stable transfectants and leukocyte cell lines expressing native PSGL-1 (HL-60 cells and Jurkat cells), and co-immunolocalization and co-immunoprecipitation of the molecules in these cells. Incubation of HL-60 cells with recombinant ADAM28s enhanced the binding to P-selectin-coated wells and P-selectin-expressing endothelial cells. In addition, intravenous injection of ADAM28s-treated HL-60 cells increased their accumulation in the pulmonary microcirculation and alveolar spaces in a mouse model of endotoxin-induced inflammation. These data suggest a novel function that ADAM28s promotes PSGL-1/P-selectin-mediated leukocyte rolling adhesion to endothelial cells and subsequent infiltration into tissue spaces through interaction with PSGL-1 on leukocytes under inflammatory conditions.
authors.affiliation: Department of Pathology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-0016, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Shimoda
authors.forename: Gakuji
authors.initials: G
authors.lastname: Hashimoto
authors.forename: Satsuki
authors.initials: S
authors.lastname: Mochizuki
authors.forename: Eiji
authors.initials: E
authors.lastname: Ikeda
authors.forename: Norihiro
authors.initials: N
authors.lastname: Nagai
authors.forename: Susumu
authors.initials: S
authors.lastname: Ishida
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Okada
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002448:Cell Adhesion / Q000235:genetics; D002522:Chlorocebus aethiops; D042783:Endothelial Cells / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000378:metabolism; D019169:Jurkat Cells; D036904:Leukocyte Rolling / Q000235:genetics; D007962:Leukocytes / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D019007:P-Selectin / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D051722:ADAM Proteins; C410224:ADAM28 protein, human