کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10804483 | 1057276 | 2005 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Local dynamics measured by hydrogen/deuterium exchange and mass spectrometry of creatine kinase digested by two proteases
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
DTTTFALC-MSMM-CKH/D - H / DMS/MS - MS / MSTrifluoroacetic acid - اسید Trifluoroacetichydrogen exchange - تبادل هیدروژنdithiothreitol - دیتیوتریتولLiquid chromatography-mass spectrometry - طیف سنجی جرم کروماتوگرافی مایعMass spectrometry - طیف سنجی جرمیTandem mass spectrometry - طیف سنجی جرمی پشت سر هم یا متوالیhydrogen/deuterium - هیدروژن / دایتریمPepsin - پپسین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله

چکیده انگلیسی
Hydrogen/deuterium exchange coupled to mass spectrometry has been used to investigate the structure and dynamics of native dimeric cytosolic muscle creatine kinase. The protein was incubated in D2O for various time. After H/D exchange and rapid quenching of the reaction, the partially deuterated protein was cleaved in parallel by two different proteases (pepsin or type XIII protease from Aspergillus saitoi) to increase the sequence coverage and spatial resolution of deuterium incorporation. The resulting peptides were analyzed by liquid chromatography coupled to mass spectrometry. In comparison with the 3D structure of MM-CK, the analysis of the two independent proteolysis deuteration patterns allowed us to get new insights into CK local dynamics as compared to a previous study using pepsin [Mazon et al. Protein Science 13 (2004) 476-486]. In particular, we obtained more information on the kinetics and extent of deuterium exchange in the N- and C-terminal extremities represented by the 1-22 and 362-380 pepsin peptides. Indeed, we observed a very different behaviour of the 1-12 and 13-22 type XIII protease peptides, and similarly for the 362-373 and 374-380 peptides. Moreover, comparison of the deuteration patterns of type XIII protease segments of the large 90-126 pepsin peptide led us to identify a small relatively dynamic region (108-114).
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimie - Volume 87, Issue 12, December 2005, Pages 1101-1110
Journal: Biochimie - Volume 87, Issue 12, December 2005, Pages 1101-1110
نویسندگان
Hortense Mazon, Olivier Marcillat, Eric Forest, Christian Vial,