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The application of rapid kinetic techniques to the transport of thymidine and 3‐O‐methylglucose into mammalian cells in suspension culture
Author(s) -
Wohlhueter Robert M.,
Marz Richard,
Graff Jon C.,
Plagemann Peter G. W.
Publication year - 1976
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040890417
Subject(s) - thymidine , nucleotide , intracellular , suspension culture , kinetics , biophysics , biochemistry , diffusion , chemistry , biology , cell culture , dna , physics , genetics , thermodynamics , gene , quantum mechanics
Techniques are described by which the transport of nutrients into mammalian cells in suspension can be measured at intervals of 1.5 seconds. By application of these techniques, the existence of a saturable (K m = 85 μM), non‐concentrative, transport system for thymidine was demonstrated in Novikoff rat hepatoma cells depleted of ATP. At concentrations of thymidine less than the K m , this system operated at velocities sufficient to nearly completely equilibrate intra‐ and extra‐cellular thymidine pools within 8 seconds. In phosphorylating cells, the transport system operated with similar rapidity, so that intracellular phosphorylation was rate‐limiting for the incorporation of thymidine into nucleotides. Uptake of 3‐O‐methylglucose occurred at comparable velocities, attaining 90% of equilibrium between internal and external pools within 25 seconds. Uptake of cytosine by simple diffusion was 100 times slower.