Kinetic lability of zinc bound to metallothionein in Ehrlich cells.
Ehrlich ascites-tumour cells normally contain a large concentration of Zn-metallothionein. When cells are placed in culture media, containing or pretreated with the metal-ion-chelating resin Chelex-100, they stop growing, remain viable and lose zinc specifically from the metallothionein (MT) pool. The kinetics of loss of zinc are first-order and are very rapid, having a rate constant of greater than or equal to 0.6 h-1. MT protein labelled with 35S is biodegraded with a rate constant of 0.07-0.014 h-1 in control cells, 0.08 h-1 in cells exposed to the zinc-deficient medium and 0.12-0.18 h-1 in cells treated directly with Chelex. Over the 6 h period in which zinc is totally lost from Zn-MT there is relatively little decrease in MT-like protein as measured by cadmium-binding to the 10,000-Mr protein fraction. Other pools of zinc and 35S-labelled protein turn over more slowly. There is no loss of zinc from rat liver Zn-MT that is dialysed against Chelex to model the possible reaction of the resin with Ehrlich-cell Zn-MT. However, Chelex does compete slowly for MT-bound zinc when resin and MT are directly mixed. Analysis of the known and possible pathways of zinc metabolism in cells in relationship to these rate constants shows that biodegradation of MT protein cannot account for the rate of loss of zinc from Zn-MT.
Krezoski
S K
SK
Department of Chemistry, University of Wisconsin-Milwaukee 53201.
Villalobos
J
J
Shaw
C F
CF
Petering
D H
DH
D000818:Animals; D002104:Cadmium / Q000378:metabolism; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism*; D002614:Chelating Agents; D002850:Chromatography, Gel; D003545:Cysteine / Q000378:metabolism; D007700:Kinetics; D008668:Metallothionein / Q000378:metabolism*; D008954:Models, Biological; D012117:Resins, Synthetic; D013462:Sulfur Radioisotopes; D014238:Trichloroacetic Acid / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
D002614:Chelating Agents; D012117:Resins, Synthetic; D013462:Sulfur Radioisotopes; D002104:Cadmium; C024997:Chelex 100; D014238:Trichloroacetic Acid; D008668:Metallothionein; D015032:Zinc; D003545:Cysteine