Fig. 2.
CD33 phosphoamino acid analysis in U937 or HL60 cells detects only phosphoserine.
[32P]-orthophosphate-labeled CD33 from PMA-stimulated U937 (A) or HL-60 cells (B) was isolated by My9 adsorption and SDS-PAGE as described in “Materials and methods.” Following hydrolysis in 6 N HCl, the resulting phosphoamino acids were mixed with phosphoserine (pS), phosphothreonine (pT), and phosphotyrosine (pY) standards and analyzed by 2-dimensional high-voltage thin-layer cellulose chromatography. Samples were applied at (+). The radioactivity above the origin represents partially digested phosphopeptides, whereas that in the upper left corner represents free phosphate.