کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9757453 | 1496004 | 2005 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Thermal unfolding of two designed monomeric λ Cro repressor variants
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کلمات کلیدی
DSCDLSOligomernuclear magnetic resonance - رزونانس مغناطیسی هستهایThermal unfolding - تابش گرماییFourier transform infrared - تبدیل فوریه مادون قرمزNMR - تشدید مغناطیسی هستهای circular dichroism - رنگ تابی دورانیFT-IR - طیف سنجی مادون قرمز تبدیل فوریهDynamic Light Scattering - پراکندگی نور دینامیکیDifferential scanning calorimetry - کالریمتری روبشی افتراقی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
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چکیده انگلیسی
The thermal unfolding of the wild-type λ Cro repressor and of two designed variants, Cro K56-[DGEVK] and Cro K56-[DGEVK] Q16L, was studied by Fourier transform infrared spectroscopy and dynamic light scattering. The engineered Cro K56-[DGEVK] monomer has five additional amino acids inserted after position 56 of the wild-type sequence, while the K56-[DGEVK] Q16L variant differs only in one position (Gln-16 to Leu substitution) from the Cro K56-[DGEVK] sequence. The temperature dependence of selected protein backbone infrared 'marker' bands revealed that Cro K56-[DGEVK] is slightly more stable than the wild-type protein, while the replacement of Gln-16 by Leu increases the thermal transition temperature by â¼20 °C. Moreover, thermal unfolding of the two Cro variants was found to proceed through equilibrium unfolding intermediates and to involve the formation of oligomers. The first thermal transition of Cro K56-[DGEVK] involves the melting of major parts of its native secondary structure and is accompanied by the formation of dimers and non-native β-sheet structures. These structures unfold during a second transition at higher temperatures, accompanied by the dissociation of the dimers. In contrast to the Cro K56-[DGEVK] protein, the intermediate state of the Cro K56-[DGEVK] Q16L variant is less well defined, and involves the formation of oligomers of different size.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Vibrational Spectroscopy - Volume 38, Issues 1â2, 29 July 2005, Pages 45-51
Journal: Vibrational Spectroscopy - Volume 38, Issues 1â2, 29 July 2005, Pages 45-51
نویسندگان
Heinz Fabian, Klaus Gast, Vladimir V. Filimonov, Dmitry F. Zamyatkin, Vladimir V. Rogov,