کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10752339 | 1050326 | 2015 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
ROZA-XL, an improved FRET based biosensor with an increased dynamic range for visualizing Zeta Associated Protein 70 kD (ZAP-70) tyrosine kinase activity in live T cells
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کلمات کلیدی
PLCγ1ZAP-70SH2SLP-76SH2 domain-containing leukocyte protein of 76 kDaFluorescence Resonance Energy Transfer (FRET) - انتقال انرژی رزونانس فلورسانس (FRET)Biosensor - بیوسنسورLAT - سالphospholipase Cγ1 - فسفولیپاز Cγ1Linker for activation of T cells - لینک دهنده برای فعال سازی سلول های TSignal transduction - هدایت سیگنالSrc Homology 2 - همبستگی Src 2T-cell receptor - گیرنده لنفوسیت T
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله

چکیده انگلیسی
Genetically encoded FRET based biosensors allow one to visualize the spatial and temporal evolution of specific enzyme activities in live cells. We have previously reported the creation of a FRET based biosensor specific for Zeta-Associated Protein â70 kD (ZAP-70) (Randriamampita et al., 2008), a Syk family protein tyrosine kinase. ZAP-70 is essential for early T cell receptor (TCR) signaling events, T lymphocyte development and has also been implicated in integrin mediated T lymphocyte migration. In order to facilitate the study of ZAP-70 kinase activity during dynamic phenomena such as immunological synapse formation or cell migration, we have designed and prepared a second generation of ZAP-70 specific biosensors. Here we describe a novel biosensor named ROZA-XL, that displays a 3-4 times greater dynamic range than its predecessor and possesses a robust baseline FRET value when expressed in the Jurkat human T cell line. We demonstrate that the robust behavior of this biosensor allows for rapid analysis of TCR mediated of ZAP-70 kinase activity at a single cell level, as shown in a simple end point assay in which ROZA-XL expressing cells are allowed to interact with stimulatory anti-CD3epsilon coated coverslips.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 459, Issue 3, 10 April 2015, Pages 405-410
Journal: Biochemical and Biophysical Research Communications - Volume 459, Issue 3, 10 April 2015, Pages 405-410
نویسندگان
Sophie Cadra, Alexia Gucciardi, Marie-Pierre Valignat, Olivier Theodoly, Aldo Vacaflores, Jon C.D. Houtman, Annemarie C. Lellouch,