Analysis of proteins binding to the ITAM motif of the beta-subunit of the high-affinity receptor for IgE (FcepsilonRI).
Aggregation of the multichain (alphabetagamma2) high-affinity IgE receptor (Fcepsilon RI) initiates a signaling cascade that results in the release of allergic mediators. The cytoplasmic tails of the FcepsilonRI-beta and -gamma subunits contain immunoreceptor tyrosine-based activation motifs (ITAMs). Phosphorylation of the gammaITAM mediates activation of Syk kinase and is sufficient for triggering the responses induced by Fcepsilon RI crosslinking. Phosphorylation of the betaITAM is insufficient to mediate cell activation. The rat betaITAM contains three tyrosines (Tyr218, Tyr224, and Tyr228) with an intermediate noncanonical tyrosine. Synthetic peptides based on the ITAM of the FcepsilonRI-beta subunit were used to investigate the role of each phosphotyrosine in the binding of signaling proteins to this motif. Among the proteins that bind to phosphorylated beta ITAM are Syk, Grb2, Shc, SHIP, and SHP-1, and binding does not depend on previous cell activation. Nonphosphorylated peptides do not bind these proteins. Syk binding to beta-peptides is dependent on the number and position of phosphotyrosines in the ITAM. Phosphorylation of Tyr218 seems to be most important for Syk binding. Recruitment of Syk and other signaling proteins to the beta-subunit might be important for its amplifier role.
Soto-Cruz
Isabel
I
Unidad de Investigación en Diferenciación Celular y Cáncer, FES, Universidad Nacional Autónoma de México, Zaragoza.
Oliver
Janet M
JM
Ortega
Enrique
E
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D017455:Receptors, IgE / Q000235:genetics / Q000378:metabolism*; D000072377:Syk Kinase; D014443:Tyrosine / Q000378:metabolism*
D047908:Intracellular Signaling Peptides and Proteins; C502447:MS4A2 protein, human; D010455:Peptides; D017455:Receptors, IgE; D014443:Tyrosine; D010770:Phosphotransferases; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C000606898:Syk protein, rat