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  • Title: Metallothionein biodegradation in rat hepatoma cells: a compartmental analysis aided 35S-radiotracer study.
    Author: Steinebach OM, Wolterbeek BT.
    Journal: Biochim Biophys Acta; 1992 Apr 22; 1116(2):155-65. PubMed ID: 1581344.
    Abstract:
    Disappearance rates, synthesis and biodegradation of rat HTC cell metallothioneins (MT) were examined by measurements of [35S]MT, and expressed by their rate constants in experiments carried out under steady-state conditions, with physiological doses of zinc and copper. Overall rates of disappearance of [35S]MT did not obey first-order kinetics, apparently due to the presence of chemically and/or spatially distinguishable MT (sub)pools. Application of compartmental analysis yielded results indicating that: (a) MT in copper-treated cells is stabler than MT in zinc-treated cells, (b) apparent half-lives for MT disappearance are 22-25 h under the conditions as described, (c) addition of 0.1 mM chloroquine reduces the overall rate of MT disappearance by 25%, (d) total MT should be regarded as significantly consisting of at least two MT sub-forms, i.e., Apo-MT and M-MT and/or cytosolic MT and lysosomal MT, each being depleted and replenished at different rates, (e) mean half-lives for total MT, based on actual degradation rates, were 3-7 h, (f) addition of 0.1 mM chloroquine resulted in both increased synthesis and increased degradation of non-lysosomal MT, the latter probably due to free amino acid depletion in the cells, and (g) MT behaviour may be best examined by combination of tracer experiments (35S-labeled amino acids), simultaneous determination of MT levels in all (sub)MT pools, and application of compartmental analysis.
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