title: Tyrosine phosphorylation of CD13 regulates inflammatory cell-cell adhesion and monocyte trafficking.
abstract: CD13 is a large cell surface peptidase expressed on the monocytes and activated endothelial cells that is important for homing to and resolving the damaged tissue at sites of injury. We showed previously that cross-linking of human monocytic CD13 with activating Abs induces strong adhesion to endothelial cells in a tyrosine kinase- and microtubule-dependent manner. In the current study, we examined the molecular mechanisms underlying these observations in vitro and in vivo. We found that cross-linking of CD13 on U937 monocytic cells induced phosphorylation of a number of proteins, including Src, FAK, and ERK, and inhibition of these abrogated CD13-dependent adhesion. We found that CD13 itself was phosphorylated in a Src-dependent manner, which was an unexpected finding because its 7-aa cytoplasmic tail was assumed to be inert. Furthermore, CD13 was constitutively associated with the scaffolding protein IQGAP1, and CD13 cross-linking induced complex formation with the actin-binding protein α-actinin, linking membrane-bound CD13 to the cytoskeleton, further supporting CD13 as an inflammatory adhesion molecule. Mechanistically, mutation of the conserved CD13 cytoplasmic tyrosine to phenylalanine abrogated adhesion; Src, FAK, and ERK phosphorylation; and cytoskeletal alterations upon Ab cross-linking. Finally, CD13 was phosphorylated in isolated murine inflammatory peritoneal exudate cells, and adoptive transfer of monocytic cell lines engineered to express the mutant CD13 were severely impaired in their ability to migrate into the inflamed peritoneum, confirming that CD13 phosphorylation is relevant to inflammatory cell trafficking in vivo. Therefore, this study identifies CD13 as a novel, direct activator of intracellular signaling pathways in pathophysiological conditions.
authors.affiliation: Center for Vascular Biology, University of Connecticut Health Center, Farmington, CT 06030.
authors.forename: Jaganathan
authors.initials: J
authors.lastname: Subramani
authors.forename: Mallika
authors.initials: M
authors.lastname: Ghosh
authors.forename: M Mamunur
authors.initials: MM
authors.lastname: Rahman
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Caromile
authors.forename: Claire
authors.initials: C
authors.lastname: Gerber
authors.forename: Karim
authors.initials: K
authors.lastname: Rezaul
authors.forename: David K
authors.initials: DK
authors.lastname: Han
authors.forename: Linda H
authors.initials: LH
authors.lastname: Shapiro
mesh_terms: D000818:Animals; D018826:CD13 Antigens / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion / Q000276:immunology; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D007249:Inflammation / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism*; D010766:Phosphorylation; D051179:Plakins / Q000378:metabolism; D011485:Protein Binding; D015398:Signal Transduction; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D051179:Plakins; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D018826:CD13 Antigens