Flow cytometric determination of cell cycle phase-specific changes in cellular phosphatase and glycosidase activities.

The activities of two phosphatases (E.C. 3.1.3.1 and 3.1.4.1) and four glycosidases (E.C. 3.2.1.21, 3.2.1.30, 3.2.1.31 and 3.2.1.51) were measured by fluorescence spectrophotometry, and flow cytometry, in mitogen-stimulated lymphocytes, and in cultures of Molt-4-F and F-89 cell lines, synchronized by hydroxyurea or thymidine. All enzymes were active throughout the cycle but the activities of three enzymes were elevated at specific points in the cycle, alkaline phosphatase activity increased at G2 + M/G1 boundary and in early S-phase, the activity of beta-L fucosidase was elevated in G1 and late S-phase. Orthophosphate diesterase activity was elevated at the G1/S boundary, and during G2 + M. The increase in beta-L fucosidase activity was due to an increased number of cells showing activity, whilst the increase in orthophosphate diesterase activity was attributable to an increase in cellular enzyme activity. Only the activities of orthophosphate diesterase and beta-L fucosidase were measurable by flow cytometry, alkaline phosphatase activity was mainly extracellular, and therefore not detectable by flow cytometric methods employed.