Treatment with okadaic acid reveals strong threonine phosphorylation of CD18 after activation of CD11/CD18 leukocyte integrins with phorbol esters or CD3 antibodies.
The CD11/CD18 leukocyte integrins comprise three heterodimers involved in leukocyte adhesion. CD11/CD18 avidity may be regulated intracellularly, and the CD18 polypeptide has previously been shown to become phosphorylated in leukocytes after phorbol ester stimulation. The importance of phosphorylation in the regulation of CD11/CD18 avidity has, however, remained unclear. We have now activated T cells using phorbol esters, CD3, and CD44 Abs. Both phorbol ester and CD3 treatment activated protein kinase C. CD18 was shown to become more stably phosphorylated after phorbol ester treatment and more transiently so after CD3 stimulation. The phosphorylation was strongly augmented by okadaic acid, a serine/threonine phosphatase inhibitor. While phorbol ester treatment caused phosphorylation mainly on serine, in okadaic acid-pretreated cells, both phorbol ester treatment as well as CD3 stimulation revealed strong threonine phosphorylation. Since earlier mutational studies have demonstrated the functional importance of cytoplasmic threonine residues in CD18, the threonine phosphorylation reported here indicates the role of threonine phosphorylation in the regulation of CD11/CD18 avidity.
Valmu
L
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Department of Biosciences, University of Helsinki, Finland.
Gahmberg
C G
CG
D000470:Alkaloids / Q000494:pharmacology; D018845:CD11 Antigens / Q000276:immunology*; D018821:CD18 Antigens / Q000276:immunology* / Q000378:metabolism*; D002449:Cell Aggregation / Q000187:drug effects; D004988:Ethers, Cyclic / Q000494:pharmacology*; D006801:Humans; D016853:Muromonab-CD3 / Q000494:pharmacology*; D019319:Okadaic Acid; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D010769:Phosphothreonine / Q000378:metabolism; D011493:Protein Kinase C / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D019311:Staurosporine; D016176:T-Lymphocyte Subsets / Q000187:drug effects* / Q000276:immunology; D013912:Threonine / Q000378:metabolism*
D000470:Alkaloids; D018845:CD11 Antigens; D018821:CD18 Antigens; D004988:Ethers, Cyclic; D016853:Muromonab-CD3; D010769:Phosphothreonine; D019319:Okadaic Acid; D013912:Threonine; D015240:Phorbol 12,13-Dibutyrate; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C; D019311:Staurosporine