کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10765112 | 1050589 | 2009 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Detection of conformationally changed MBP using intramolecular FRET
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کلمات کلیدی
MBPIPTGEYFPECFPSDS–PAGE - SDS-PAGEenhanced cyan fluorescent protein - افزایش پروتئین فلورسنت سیانوژنsodium dodecyl sulfate–polyacrylamide gel electrophoresis - الکتروفورز ژل دوده سولفات سدیم پلی آکریل آمیدFluorescence resonance energy transfer - انتقال انرژی رزونانس FluorescenceFRET - انتقال انرژی رزونانسی فورسترisopropyl β-D-thiogalactopyranoside - ایزوپروپیل β-D-thiogalactopyranosideConformational change - تغییر کنفورماسیونیenhanced yellow fluorescent protein - پروتئین فلورسنت زرد افزایش یافته استmaltose binding protein - پروتئین متصل به مالتوز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله

چکیده انگلیسی
The principal objective of this study was to explore protein conformational changes using fluorescence resonance energy transfer (FRET) technology. Maltose binding protein (MBP) was adopted as a target model, due to its well-characterized structure and ligand specificity. To the best of our knowledge, this is the first report to provide information regarding the biological distance between the two lobes of MBP upon maltose binding. For the FRET pair, ECFP and EYFP were used as the donor and the acceptor, and were linked genetically to the C-terminal and N-terminal regions of MBP (ECFP:MBP:EYFP), respectively. After the FRET reaction, maltose-treated MBP was shown to exhibit a considerable energy transfer (FRET efficiency (E)Â =Â â¼0.11, Distance (D)Â =Â â¼6.93Â nm) at the ensemble level, which was regarded as reflective of the increase in donor quenching and the upshift in acceptor emission intensity, thereby suggesting that the donor and the acceptor had been brought close together as the result of structural alterations in MBP. However, upon glucose treatment, no FRET phenomenon was detected, thereby implying the specificity of interaction between MBP and maltose. The in vitro FRET results were also confirmed via the acceptor photobleaching method. Therefore, our data showed that maltose-stimulated conformational changes of MBP could be measured by FRET, thereby providing biological information, including the FRET efficiency and the intramolecular distance.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 388, Issue 3, 23 October 2009, Pages 560-564
Journal: Biochemical and Biophysical Research Communications - Volume 388, Issue 3, 23 October 2009, Pages 560-564
نویسندگان
Kyoungsook Park, Lan Hee Lee, Yong-Beom Shin, So Yeon Yi, Yong-Won Kang, Dai-Eun Sok, Jin Woong Chung, Bong Hyun Chung, Moonil Kim,