Effects of methyl mercuric chloride and sulfhydryl inhibitors on phospholipid synthetic activity of lymphocytes
✍ Scribed by Katsuhiro Kageyama; Yasuto Onoyama; Eiichi Kano
- Publisher
- John Wiley and Sons
- Year
- 1986
- Tongue
- English
- Weight
- 458 KB
- Volume
- 6
- Category
- Article
- ISSN
- 0260-437X
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✦ Synopsis
The effect of methyl mercuric chloride on the activity of phospholipid synthesis of rat lymph node lymphocytes was compared with that of sulfhydryl inhibitors. Measurement of the radioactivity of [ ''C]oleic acid and [ 14C]acetate incorporated into lecithin of cells during short-term incubation showed that all the inhibitors tested similarly reduced the incorporation. However, methyl mercuric chloride (MMC) was the strongest inhibitor, being effective at 4 pM and causing more than 80% decrease at 20 p ~.
Inhibition by the sulfhydryl inhibitors, at less than 40 p M , ranked as follows: N-ethylmaleimide > a-chloroacetophenone > hydroquinone > iodoacetamide. MMC also obstructed the enhancement by phytohemagglutinin of ['4C]oleic acid incorporation into lecithin. MMC was effective at 2 p M , while the other agents had little or no effect at this concentration. Further investigation suggested that inhibition of phospholipid synthesis did not depend on reduced incorporation of oleic acid into the cellular membrane but on decreased turnover of the fatty acid into phospholipids after the incorporation. The viability of lymphocytes incubated with the agents was measured by trypan blue dye-exclusion test. More than 90% of the cells treated with MMC at a concentration as low as 20-40pM died, but the SH inhibitors, including NEM which greatly inhibited the phospholipid synthesis, produced few cell deaths at these concentrations.
These observations show that the SH inhibitors affect enzymes in phospholipid synthesis, whereas MMC not only inhibits the enzymes but kills cells.
EXPERIMENTAL
Agent and substrate solutions MMC (Tokyo Kasei Kogyo Co. Ltd, Japan) and NEM, achloroacetophenone (a-CAP), IAA and hydroquinone (HQ)