کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1177546 962026 2006 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A biosensor-based approach toward purification and crystallization of G protein-coupled receptors
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
A biosensor-based approach toward purification and crystallization of G protein-coupled receptors
چکیده انگلیسی

Biacore technology was used to develop an affinity purification method and screen cocrystallization conditions for the chemokine receptor CCR5. We characterized the binding of nine HIV gp120 variants and identified a truncated construct (YU2DV1V2) that bound CCR5 independent of CD4. This construct was used in an affinity purification step to improve the activity of detergent-solubilized receptor by approximately 300%. The biosensor was also used to screen receptor binding activity automatically under 50 different crystallization conditions. We found that high-molecular-weight polyethylene glycols (PEGs 4000 and 8000 Da) most often stabilized the receptor and improved complex formation with potential cocrystallization partners such as conformationally sensitive monoclonal antibodies and gp120. Our results show how biosensors can provide unique insights into receptor purification methods and reveal the effects of crystallization conditions on complex formation. Importantly, these methods can be readily applied to other systems.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 353, Issue 2, 15 June 2006, Pages 278–283
نویسندگان
, , , ,