کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1926392 1536463 2009 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Identifying calpain substrates in intact S2 cells of Drosophila
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Identifying calpain substrates in intact S2 cells of Drosophila
چکیده انگلیسی

Calpains are cysteine proteases involved in a number of physiological and pathological processes, yet our knowledge of substrates cleaved in vivo, in intact cells, is scarce. In this work we made an attempt to develop a technique for finding calpain substrates in intact Drosophila Schneider S2 cells. The procedure consists in comparative 2D gelelectrophoresis: three identical samples were treated in different ways: A (control, no addition), B, activated (Ca2+ and ionomycin added), C, inactivated (additions as in B + specific calpain inhibitor). 2D gel pattern were analyzed by densitometry. Spots showing density relation A > B << C were identified by mass spectroscopy. In a typical run, 11 candidate substrates were recognized; out of these, four were randomly selected: all four were verified to be calpain substrates, by digestion of the recombinant protein with recombinant calpain.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Archives of Biochemistry and Biophysics - Volume 481, Issue 2, 15 January 2009, Pages 219–225
نویسندگان
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