کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1173944 1491400 2014 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Weak protein–protein interactions revealed by immiscible filtration assisted by surface tension
ترجمه فارسی عنوان
تداخل پروتئین های ضعیف با ترکیبات نیتروژن با استفاده از تنش سطحی به دست می آید
کلمات کلیدی
ایمن سازی، پاسخ پالئول، پروتئین فلورسنت سبز، گیرنده لیپوپروتئین با چگالی کم، گلیکوزین سیتستاز کیناز 3β، آماده سازی نمونه بر مبنای محرومیت
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی

Biological mechanisms are often mediated by transient interactions between multiple proteins. The isolation of intact protein complexes is essential to understanding biochemical processes and an important prerequisite for identifying new drug targets and biomarkers. However, low-affinity interactions are often difficult to detect. Here, we use a newly described method called immiscible filtration assisted by surface tension (IFAST) to isolate proteins under defined binding conditions. This method, which gives a near-instantaneous isolation, enables significantly higher recovery of transient complexes compared to current wash-based protocols, which require reequilibration at each of several wash steps, resulting in protein loss. The method moves proteins, or protein complexes, captured on a solid phase through one or more immiscible-phase barriers that efficiently exclude the passage of nonspecific material in a single operation. We use a previously described polyol-responsive monoclonal antibody to investigate the potential of this new method to study protein binding. In addition, difficult-to-isolate complexes involving the biologically and clinically important Wnt signaling pathway were isolated. We anticipate that this simple, rapid method to isolate intact, transient complexes will enable the discoveries of new signaling pathways, biomarkers, and drug targets.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 447, 15 February 2014, Pages 133–140
نویسندگان
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