EFFECT OF POLYAMINE ANALOGS ON HYPUSINE CONTENT IN JURKAT T-CELLS

The availability of synthetic hypusine and deoxyhypusine has made it p ossible to develop analytical methods which allow for the measurement of these compounds in various tissues. The methods involve dansylation of extracts fi om the pellet remaining after perchloric acid precipit ation of cell or tissue homogenates, followed by high-performance liqu id chromatography. To demonstrate the utility of this approach, the im pact of four polyamine analogues, N-1,N-11- diethylnorspermine (DENSPM ), N-1,N-14-diethylhomospermine (DEHSPM), 1,6,12-triazadodecane [(4,5) triamine], and 1,7,13-triazatridecane [(5,5) triamine], on hypusine l evels in a human T-cell line (JURKAT) is evaluated. All four analogues are active in controlling cell growth and compete well with spermidin e for the polyamine transport apparatus. After 144 h of exposure to JU RKAT cells, DENSPM reduces putrescine to below detectable limits and s permidine to 10% of the level in control cells. The other three analog ues diminish both putrescine and spermidine to below detectable limits . The effectiveness with which the compounds lower spermine levels is DENSPM > DEHSPM > (4,5) triamine > (5,5) triamine. The analogues decre ase the activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase in a similar fashion. Of the four polyamines, DENSPM an d DEHSPM are potent at lowering intracellular hypusine levels after 14 4 h: 59 +/- 9% and 73 +/- 12% of control levels, respectively. The oth er two analogues have marginal effects.