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					Apyrase inhibitors affect pollen growth


       Discovery Lab Lecture
         February 7, 2008
Apyrase inhibitors were discovered in a
screen of 9,000 small organic molecules
   Assay tested whether randomly selected
    organic compounds could block the ability of
    potato apyrase to remove phosphates from
    ATP
   Only 17 compounds were found to be
    effective inhibitors
   These inhibitors differed in their relative
    specificity
   These were the first compounds found to be
    very effective inhibitors of apyrase activity
    Inhibitors NGXT 191, NGXT 1913 and #4
differ in their ability to inhibit other phosphatases

Table S1. Relative Inhibitory Activity of Apyrase Inhibitors 1, 4, and 13 toward Potato
                                                       1
Apyrase , Acid Phosphatase and Alkaline Phosphatases

Inhibitor               Potato Apyrase          Acid Phosphatase       Alkaline Phosphatase
NGXT 191                     89                         0                     83
NGXT 1913                  100                          8                     27
       # 4                   73                        86                     15
1
  Numbers given in % inhibition. Data taken from Windsor, B. (2000). Establishing a role for
ecto-phosphatase in drug resistance. Ph.D. dissertation, The University of Texas at Austin.
Details of assay are given in the dissertation and in: Windsor, J.B., Thomas, C., Hurley, L., S. J.
Roux, S.J., and Lloyd, A.M. (2002). An automated colorimetric screen for apyrase inhibitors.
Biotechniques 33, 1024-1030.
NGXT 191 and Inhibitor #4 simultaneously
inhibit pollen tube growth and raise the [eATP]
Sample results from last week
   Low: 0.32 vs 0.46; 0.03 vs 0.12;
             0.36 vs. 0.15; 2.21 vs. 0.56

    High: 0.14 vs. 0.15; 0.15 vs. 0.15;
          0.08 vs. 0.07; 0.07 vs. 0.01
    This week:
 Review data from last week √
 Training on how to make bar graphs (for next week’s

  presentations and reports).
 Transfer samples planted last week to test media

  (Round 1 of experiments):
             > low (30 mM) or high (150 mM) GTPS
             > inhibitor #2 (various concentrations)
             > DPI (5 or 10 mM) + 30 mM ADPS
Then measure effects on root hair growth
Next week:
   Those giving oral reports need to load their
    presentations between 2:00 and 2:25 PM on
    Thursday (or send file to Dr. Clark by e-mail &
    ask him to load it). NOTE: If you have new
    Vista operating system, save your .ppt in old
    system.
   Those giving written reports, send your file to
    Dr. Clark by 2:15 on Thursday, and bring a
    hard copy to class.
Next week

				
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posted:8/27/2012
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