کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2065541 1076927 2010 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Recombinant expression in human cells of active integrin α1β1-blocking RTS-disintegrin jerdostatin
کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوشیمی، ژنتیک و زیست شناسی مولکولی (عمومی)
پیش نمایش صفحه اول مقاله
Recombinant expression in human cells of active integrin α1β1-blocking RTS-disintegrin jerdostatin
چکیده انگلیسی

Jerdostatin, an RTS short disintegrin cloned from Protobothrops jerdonii and recombinantly produced in Escherichia coli, is a potent and specific antagonist of the α1β1 integrin. Jerdostatin selectively blocked the adhesion of α1β1-K562 cell to collagens I and IV in vitro and angiogenesis in vivo. Here we report the recombinant production of jerdostatin in a mammalian cell system, a prerequisite for developing a conditional transgenic mouse to investigate the effect of systemic expression of jerdostatin on tumor development. For proper export of jerdostatin, a secretion leader sequence was engineered at the protein’s N-terminus. A FLAG epitope was also included at the N-terminus of the mature disintegrin to facilitate its isolation and characterization of recombinant jerdostatin (rJerd). This pRc-CMV/FLAG-rJerd construct was transiently expressed in HEK-293 cells and was efficiently secreted into the culture medium. rJerd bound to recombinant soluble α1β1 integrin in a saturable and cation-independent manner. Soluble rJerd also inhibited the binding of α1β1 integrin to the CB3 fragment of collagen IV in a dose-dependent manner (IC50 570 nM). Mammalian cell-expressed jerdostatin disrupted the adhesion of RuGli cells to collagen IV. Our results highlight pRc-CMV/FLAG-rJerd as a suitable construct for expressing soluble active α1β1-blocking jerdostatin in a mammalian cell system.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Toxicon - Volume 56, Issue 6, November 2010, Pages 1052–1058
نویسندگان
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