Sykis phosphorylated on multiple tyrosine sites, including Tyr317, Tyr342, and Tyr346 in interdomain B under the condition of binding with B-cell receptor , and phosphorylated on Tyr630 of human Syk to create binding site for BLNK .
Chan, "BLNK: a central linker protein in B cell activation," Immunity, vol.
However in T cells we have found that attenuation of ERK by [Hg.sup.2+] is associated with attenuation of ZAP-70, as well as LAT, the scaffolding protein homolog of BLNK .
Analysis on the single cell level shows that while activation of ERK is suppressed, upstream activation of Syk is enhanced and phosphorylation of BLNK is increased.
Low Levels of [Hg.sup.2+] Attenuate Phosphorylation of ERK, Accentuate Phosphorylation of Syk and BLNK, but Has Little Effect on Phosphorylation of BTK during BCR Mediated Signal Transduction in WEHI-231 Cells.
The cells were resuspended in an antibody cocktail containing fluorescently labeled antibodies specific for the phosphorylated residues of ERK1/2, Syk, BLNK, or Btk according to the manufacturer's instructions.
Fluorescently labeled antibodies used for cytometric analysis of phosphorylated ERK1/2, Syk, BLNK, and Btk were purchased from BD Biosciences (San Jose, CA).
In Figure 4 we reduced complex raw data sets obtained from the flow cytometery so as to characterize different cell populations solely with four numbers, the four MFIs associated with binding phosphospecific antibodies to ERK, Syk, BTK and BLNK. The effect of [Hg.sup.2+] on BCR signal transduction was then essentially determined by comparing these numbers among differentially treated cell populations.