ARACHIDONOYL AMINO ACIDS AND ARACHIDONOYL PEPTIDES
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washed pellet was resuspended in a 10 mM Tris-HCl
buffer (pH 7.4) containing 0.22 M saccharose.
A mixture of [3H8]AA-MEHFPGP solution in
buffer B (final volume 200 µl) with a membrane sus-
pension (protein concentration 0.2 mg/ml) in buffer B
containing 1 mM benzamidine and 0.1 mM PMSF was
incubated at 30°C for 30 min under stirring. The sam-
ples were then filtered through plate filters previously
soaked in the buffer B for 2 h. The filters were thrice
washed with buffer B (200 µl). After drying the plates
on air, the filters were removed and transferred into
scintillation vials containing a scintillation mixture (4
ml). The amount of label retained in the membranes in
the presence of 0.1 mM unlabeled peptide was taken as
nonspecific binding.
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For assaying the stability of arachidonoyl-Semax,
the peptide (10 nmol) was incubated with the mem-
branes as described above without addition of inhibi-
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ACKNOWLEDGMENTS
17. Austin-Brown, S.L. and Chapman, K.D., Plant Physiol.,
2002, vol. 129, pp. 1892–1898.
This work was partly supported by the Russian
Foundation for Basic Research (project nos.
04-04-81028-Bel2004_a, 05-04-08132-ofi_a, and
05-04-48293-a), the President Program for Support of
Leading Scientific Schools of Russia (project no.
NSh-2150.2003.4 under the direction of academician
N.F. Myasoedov), and the Belarussian Republican
Foundation for Basic Research (project no. B04R-135).
18. Burstein, S.H., Rossetti, R.G.,Yagen, B., and Zurier, R.B.,
Prostaglandins Other Lipid Mediat., 2000, vol. 61,
pp. 29–41.
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