کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5507006 1536898 2017 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
The effect of alcohol on recombinant proteins derived from mammalian adenylyl cyclase
ترجمه فارسی عنوان
اثر الکل بر پروتئین های نوترکیب حاصل از آدنیلیل سیکلاز پستانداران
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
چکیده انگلیسی


- The activity of catalytic domains of mammalian adenylyl cyclase respond to ethanol in an isoform specific manner.
- The alcohol cutoff effect of the recombinant adenylyl cyclase type 7 is same as that observed for the native protein.
- Alcohol appears to interact with adenylyl cyclase type 7 at multiple sites with positive cooperativity.

The cyclic AMP (cAMP) signaling pathway is implicated in the development of alcohol use disorder. Previous studies have demonstrated that ethanol enhances the activity of adenylyl cyclase (AC) in an isoform specific manner; AC7 is most enhanced by ethanol, and regions responsible for enhancement by ethanol are located in the cytoplasmic domains of the AC7 protein. We hypothesize that ethanol modulates AC activity by directly interacting with the protein and that ethanol effects on AC can be studied using recombinant AC in vitro. AC recombinant proteins containing only the C1a or C2 domains of AC7 and AC9 individually were expressed in bacteria, and purified. The purified recombinant AC proteins retained enzymatic activity and isoform specific alcohol responsiveness. The combination of the C1a or C2 domains of AC7 maintained the same alcohol cutoff point as full-length AC7. We also find that the recombinant AC7 responds to alcohol differently in the presence of different combinations of activators including MnCl2, forskolin, and Gsα. Through a series of concentration-response experiments and curve fitting, the values for maximum activities, Hill coefficients, and EC50 were determined in the absence and presence of butanol as a surrogate of ethanol. The results suggest that alcohol modulates AC activity by directly interacting with the AC protein and that the alcohol interaction with the AC protein occurs at multiple sites with positive cooperativity. This study indicates that the recombinant AC proteins expressed in bacteria can provide a useful model system to investigate the mechanism of alcohol action on their activity.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemistry and Biophysics Reports - Volume 10, July 2017, Pages 157-164
نویسندگان
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