[Microspectrometric quantification of 4-hydroxypentenal binding to protein thiols in Ehrlich ascites tumor cells]

Abstract

After 30 minutes in 5.10(-3) M hydroxypentenal (HPE) the microscopically determinable proteinthiols (PSH) stained with DDD-Fastblue B decrease from 1,07 . 10(-14) to 0,61 . 10(-14) moles/cell. The loss of PSH of 0,46 . 10(-14) moles has a confidential range (99%) of +/- 0,12. As HPE reacts with thiols by an addition to the C3 = C2-group, the aldehydic group remains reactive and can be recognized intracellulary and quantitatively determined with 2,4-dinitrophenylhydrazine, whereby an uptake of 0,63 . 10(-14) moles of HPE per single cell is observed (confidential range +/- 0,11). The broad overlapping of the confidential ranges indicates clearly that the loss of PSH corresponds to the uptake of HPE. From the statistical distribution diagrams it can be seen that the PSH of practically all cells take part in the reaction, if however, to very different extent; the cells with high PSH concentrations being the most reactive ones.