The kinetic dissection of transport from metabolic trapping during substrate uptake by intact cells. Uridine uptake by quiescent and serum-activated nil 8 hamster cells and their murine sarcoma virus-transformed counterparts

Ora Heichal, Dvorah Ish-Shalom, Ruth Koren, Wilfred D. Stein

Research output: Contribution to journalArticlepeer-review

Abstract

1. 1. We present a theoretical analysis of the tandem processes of transport and metabolic trapping which together constitute uptake of a substrate by intact cells. 2. 2. Transport is assumed to occur by means of a simple carrier here analysed in its general form. Trapping is assumed to occur by a simple enzymic reaction. 3. 3. We show how to obtain the separate parameters of the two steps by analysing uptake data over a range of uptake times and substrate concentrations. 4. 4. We present uptake data for uridine and cytosine-β-d-arabinoside entering Nil 8 hamster fibroblasts, normal and murine sarcoma virus transformed, in the quiescent condition and after stimulation by added serum. We analyse the data in terms of the theory for tandem processes. 5. 5. Transport is characterised by a system having a high Km and a high V for entry. The data for cytosine-β-d-arabinoside suggest that the cytosine-β-d-arabinoside system is not far from a symmetric one. The data for uridine transport do not differ when quiescent serum-activated cells are compared. Transformed cells transport uridine at half the maximum velocity of normal cells, with or without added serum. 6. 6. Trapping of cytosine-β-d-arabinoside is insignificant. Trapping of uridine occurs by a system with both V and Km at least an order of magnitude smaller than are these parameters for transport. Trapping of uridine by non-transformed cells activated by serum, has twice the V of such cells in the quiescent state. 7. 7. In the virus-transformed cells, the control of uridine trapping by added serum lost, along with control of growth by this statement.

Original languageEnglish
Pages (from-to)169-186
Number of pages18
JournalBBA - Biomembranes
Volume551
Issue number1
DOIs
StatePublished - 20 Feb 1979
Externally publishedYes

Keywords

  • (Nil 8 hamster cell)
  • Transformation
  • Transport kinetics
  • Uridine uptake

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