Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10771264 | Biochemical and Biophysical Research Communications | 2005 | 5 Pages |
Abstract
1-Butanol is commonly used as a substrate for phospholipase D (PLD) activity measurement. Surprisingly we found that, in the presence of 30 mM 1-butanol (standard PLD assay conditions), PLD1 activity in COS-7 cells was lost after incubation for 2 min. In contrast, in the presence of the protein kinase C (PKC) inhibitor staurosporine or dominant negative PKCα D481E, the activity was sustained for at least 30 min. The binding between PLD1 and PKCα was also lost after 2 min incubation with 30 mM 1-butanol while staurosporine and D481E maintained the binding. 1-Butanol at 2 mM did not inhibit PLD1 basal activity or PLD1 binding to PKCα, and staurosporine and PKCα D481E produced a constant increase in PLD1 basal activity of 2-fold. These results indicate that 1-butanol is inhibitory to PLD1 activity by reducing its association with PKCα, and that the concentration of 1-butanol is an important consideration in assaying basal PLD1 activity.
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Authors
Tianhui Hu, John H. Exton,