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Flow cytometric determination of cell cycle phase‐specific changes in cellular phosphatase and glycosidase activities
Author(s) -
Britten R. A.,
Dyson J. E. D.
Publication year - 1987
Publication title -
cell biochemistry and function
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.933
H-Index - 61
eISSN - 1099-0844
pISSN - 0263-6484
DOI - 10.1002/cbf.290050103
Subject(s) - flow cytometry , alkaline phosphatase , enzyme , phosphatase , enzyme assay , cell cycle , biochemistry , chemistry , extracellular , microbiology and biotechnology , biology , cell
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 mitogenstimulated lymphocytes, and in cultures of Molt‐4‐F and F‐89 cell lines, syncronized 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 G 2 + M/G 1 boundary and in early S‐phase, the activity of β‐ L fucosidase was elevated in G 1 and late S‐phase. Orthophosphate diesterase activity was elevated at the G 1 /S boundary, and during G 2 + M. The increase in β‐ 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 β‐ L fucosidase were measurable by flow cytometry, alkaline phosphatase activity was mainly extracellular, and therefore not detectable by flow cytometric methods employed.