The cytoskeletal association of CD11/CD18 leukocyte integrins in phorbol ester-activated cells correlates with CD18 phosphorylation.
Leukocyte adhesion is a regulated process, which involves CD11/CD18 leukocyte integrins. CD11/CD18 acidity may be regulated intracellularly, and the CD18 polypeptide has previously been shown to become phosphorylated on serine and threonine after phorbol ester activation of T cells. Increased adhesiveness is believed to be mediated by regulating the overall avidity of cellular contact. CD11/CD18 integrins have earlier been reported to interact with several cytoskeletal proteins. We have now studied the involvement of the CD18 phosphorylation in cytoskeletal associations. We have investigated the distribution of phosphorylated CD18 between soluble, cytoskeletal and nuclear fractions of T cell detergent lysates. A significant amount of phosphorylated CD18 polypeptides was observed to fraction along with the cytoskeleton, while the majority of the cell surface CD18 molecules remained in the soluble fraction. Putative candidates for this altered cytoskeletal binding of CD11/CD18 were shown to be talin and filamin, which were observed to bind to CD18 cytoplasmic peptides and co-precipitate with CD18. The importance of the CD18 cytoplasmic domain in the regulation of the leukocyte adhesion was further strengthened by inhibition of phorbol ester-induced T cell adhesion with a phosphorylated lipopeptide corresponding to the cytoplasmic portion of the CD18. These results indicate that the induced CD18 phosphorylation and the altered cytoskeletal binding of the phosphorylated integrin complex may contribute to the increased avidity of CD11/CD18-mediated leukocyte adhesion.
Valmu
L
L
Department of Biosciences, University of Helsinki, Finland.
Fagerholm
S
S
Suila
H
H
Gahmberg
C G
CG
D000595:Amino Acid Sequence; D018845:CD11 Antigens / Q000378:metabolism*; D018821:CD18 Antigens / Q000737:chemistry / Q000378:metabolism*; D002448:Cell Adhesion; D003285:Contractile Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000276:immunology*; D064448:Filamins; D006801:Humans; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D008840:Microfilament Proteins / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D010766:Phosphorylation; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000502:physiology; D016608:Talin / Q000378:metabolism
D018845:CD11 Antigens; D018821:CD18 Antigens; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D016608:Talin; D019319:Okadaic Acid; D015240:Phorbol 12,13-Dibutyrate