کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1936570 | 1050695 | 2008 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Femtosecond spectroscopy probes the folding quality of antibody fragments expressed as GFP fusions in the cytoplasm
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کلمات کلیدی
Single chain variable fragment (scFv)Protein folding - تاشدگی پروتئینprotein aggregation - تجمع پروتئینExcited-state dynamics - دینامیک دولت هیجان زدهFemtosecond spectroscopy - طیف سنجی فموتوسجدFluorescence - فلورسنسGenetic fusion - همجوشی ژنتیکیGreen fluorescent protein (GFP) - پروتئین فلورسنت سبز (GFP)
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Time-resolved femtosecond spectroscopy can improve the application of green fluorescent proteins (GFPs) as protein-folding reporters. The study of ultrafast excited-state dynamics (ESD) of GFP fused to single chain variable fragment (scFv) antibody fragments, allowed us to define and measure an empirical parameter that only depends on the folding quality (FQ) of the fusion. This method has been applied to the analysis of genetic fusions expressed in the bacterial cytoplasm and allowed us to distinguish folded and thus functional antibody fragments (high FQ) with respect to misfolded antibody fragments. Moreover, these findings were strongly correlated to the behavior of the same scFvs expressed in animal cells. This method is based on the sensitivity of the ESD to the modifications in the tertiary structure of the GFP induced by the aggregation state of the fusion partner. This approach may be applicable to the study of the FQ of polypeptides over-expressed under reducing conditions.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 366, Issue 4, 22 February 2008, Pages 878-884
Journal: Biochemical and Biophysical Research Communications - Volume 366, Issue 4, 22 February 2008, Pages 878-884
نویسندگان
P. Didier, E. Weiss, A.-P. Sibler, P. Philibert, P. Martineau, J.-Y. Bigot, L. Guidoni,