کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8514303 | 1556506 | 2017 | 31 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Impact of Mutations on the Higher Order Structure and Activity of a Recombinant Uricase
ترجمه فارسی عنوان
اثر موتاسیون بر ساختار نظم بالاتر و فعالیت اوریکان بازآفرین
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کلمات کلیدی
DSCDLSSEC-MALSsize-exclusion chromatography with multi-angle light scatteringEnzymes - آنزیمUric acid - اسید اوریکBiotechnology - بیوتکنولوژیmutation - جهشbiopharmaceutical characterization - خصوصیات بیوفرمارکتیوProtein structure - ساختار پروتئینDifferential scanning fluorimetry - فلوریمتری اسکن دیفرانسیلDynamic Light Scattering - پراکندگی نور دینامیکیDifferential scanning calorimetry - کالریمتری روبشی افتراقی
موضوعات مرتبط
علوم پزشکی و سلامت
داروسازی، سم شناسی و علوم دارویی
اکتشاف دارویی
چکیده انگلیسی
This study explores the structural and functional changes associated with a low-temperature thermal transition of 2 engineered bacterial uricase mutants. Uricase has a noncovalent homotetrameric structure, with 4 active sites located at the interface of subunits. Using differential scanning calorimetry, a low-temperature transition was identified at 42°C for mutant A and at 33°C for mutant B. This transition was stabilized by the uricase inhibitor, oxonic acid, suggesting a strong structural relationship to the active site. For mutant B, there was a reversible loss of enzymatic activity above the low-temperature transition. Spectroscopic measurements demonstrated that there was also a reversible loss of secondary and tertiary structures and an increase in surface hydrophobicity. However, the hydrophobic core environment and the tetrameric structure were not altered over the low-temperature transition suggesting that the changes occurred primarily at the surface of the enzyme. The protein became aggregation-prone at temperatures approaching the cluster of higher-temperature melting transitions at 84°C, indicating these transitions represent a global unfolding of the protein. Our findings shed light on the structural changes that affect the uricase mechanism of action and provide new insights into how enzyme therapeutic development may be approached.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmaceutical Sciences - Volume 106, Issue 4, April 2017, Pages 1018-1024
Journal: Journal of Pharmaceutical Sciences - Volume 106, Issue 4, April 2017, Pages 1018-1024
نویسندگان
Flaviu Gruia, Arun Parupudi, Manuel Baca, Chris Ward, Andrew Nyborg, Richard L. Jr., Jared S. Bee,