Polarization and interaction of adhesion molecules P-selectin glycoprotein ligand 1 and intercellular adhesion molecule 3 with moesin and ezrin in myeloid cells.
In response to the chemoattractants interleukin 8, C5a, N-formyl-methionyl-leucyl-phenylalanine, and interleukin 15, adhesion molecules P-selectin glycoprotein ligand 1 (PSGL-1), intercellular adhesion molecule 3 (ICAM-3), CD43, and CD44 are redistributed to a newly formed uropod in human neutrophils. The adhesion molecules PSGL-1 and ICAM-3 were found to colocalize with the cytoskeletal protein moesin in the uropod of stimulated neutrophils. Interaction of PSGL-1 with moesin was shown in HL-60 cell lysates by isolating a complex with glutathione S-transferase fusions of the cytoplasmic domain of PSGL-1. Bands of 78- and 81-kd were identified as moesin and ezrin by Western blot analysis. ICAM-3 and moesin also coeluted from neutrophil lysates with an anti-ICAM-3 immunoaffinity assay. Direct interaction of the cytoplasmic domains of ICAM-3 and PSGL-1 with the amino-terminal domain of recombinant moesin was demonstrated by protein-protein binding assays. These results suggest that the redistribution of PSGL-1 and its association with intracellular molecules, including the ezrin-radixin-moesin actin-binding proteins, regulate functions mediated by PSGL-1 in leukocytes stimulated by chemoattractants.
Alonso-Lebrero
J L
JL
Servicio de Inmunología, Hospital de la Princesa, Madrid, Spain; Northwestern University Medical School, Chicago, IL, USA.
Serrador
J M
JM
Domínguez-Jiménez
C
C
Barreiro
O
O
Luque
A
A
del Pozo
M A
MA
Snapp
K
K
Kansas
G
G
Schwartz-Albiez
R
R
Furthmayr
H
H
Lozano
F
F
Sánchez-Madrid
F
F
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000097:blood*; D002462:Cell Membrane / Q000737:chemistry; D002846:Chromatography, Affinity; D015936:Complement C5a / Q000494:pharmacology; D003593:Cytoplasm / Q000737:chemistry; D003598:Cytoskeletal Proteins; D006801:Humans; D007118:Immunoassay; D016209:Interleukin-8 / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000097:blood*; D008840:Microfilament Proteins / Q000097:blood*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000737:chemistry* / Q000648:ultrastructure; D010750:Phosphoproteins / Q000097:blood*; D011994:Recombinant Proteins / Q000097:blood; D000099149:Ezrin
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D015815:Cell Adhesion Molecules; D015936:Complement C5a; D003598:Cytoskeletal Proteins; D016209:Interleukin-8; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D009240:N-Formylmethionine Leucyl-Phenylalanine; D010750:Phosphoproteins; D011994:Recombinant Proteins; D000099149:Ezrin; C483616:ICAM3 protein, human; C084882:P-selectin ligand protein; C071058:moesin