کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
7231825 1470957 2015 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Micro-structured peptide surfaces for the detection of high-affinity peptide-receptor interactions in living cells
ترجمه فارسی عنوان
سطوح پپتید میکرو ساختار یافته برای تشخیص اثر متقابل گیرنده های پپتیدی باکتری در سلول های زنده
کلمات کلیدی
عکسبرداری، آزمایش زنده سلولی، استرپتاویدین میکرو ساختار یافته، میکرو الگوی، چسبندگی سلولی، تعامل گیرنده پپتیده،
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی
Peptide ligands have great potential as selective agents for diagnostic imaging and therapeutic targeting of human cancers. A number of high-throughput assays for screening potential candidate peptides have been developed. Although these screening assays are indispensable for the identification of peptide ligands at a large scale, it is crucial to validate peptide binding and selectivity for targeted receptors in a live-cell context. For testing high-affinity peptide-receptor interactions in the plasma membrane of living cells, we developed cell-resistant, micro-structured glass surfaces with high-density and high-contrast peptide features. Cell adhesion and recruitment of fluorescent receptors to micro-patterned peptides in the live-cell membrane were evaluated by reflection interference contrast (RIC) and total internal reflection (TIRF) microscopy, respectively. To demonstrate both the specificity and modularity of the assay, co-patterning of fluorescent receptors with three different immobilized micro-structured ligands was shown: first, interaction of green fluorescent protein (GFP)-tagged epidermal growth factor (EGF) receptor expressed in Jurkat cells with immobilized EGF was detected and quantified. Second, using Jurkat cells, we demonstrated specific interaction of yellow fluorescent protein (YFP)-tagged β3 integrin with c(RGDfK) peptide. Third, we identified indirect recruitment of GFP-tagged α5 integrin to an 11-mer peptide. In summary, our results show that the developed micro-structured surfaces are a useful tool for the validation and quantification of peptide-receptor interactions in their natural cellular environment.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 74, 15 December 2015, Pages 757-763
نویسندگان
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