کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10798110 | 1053295 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Large-scale purification of the proton pumping pyrophosphatase from Thermotoga maritima: A “Hot-Solve” method for isolation of recombinant thermophilic membrane proteins
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کلمات کلیدی
NTADTTPPIPMSFDDMTVPSelf-organizing mappingTRISH+-PPase - H + -PPasen-dodecyl-β-d-maltopyranoside - n-dodecyl-β-D-maltopyranosideSOM - WHOnitriloacetic acid - اسید نیتریلواتیکSingle particle analysis - تجزیه و تحلیل ذرات تکThermotoga maritima - ترموتوگا maritimaDetergent solubilization - محلول سازی مواد شویندهMeS - مسElectron microscopy - میکروسکوپ الکترونیinorganic pyrophosphate - پیرو فسفات معدنی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Although several proton-pumping pyrophosphatases (H+-PPases) have been overexpressed in heterologous systems, purification of these recombinant integral membrane proteins in large amounts in order to study their structure-function relationships has proven to be a very difficult task. In this study we report a new method for large-scale production of pure and stable thermophilic H+-PPase from Thermotoga maritima. Following overexpression in yeast, a “Hot-Solve” procedure based on high-temperature solubilization and metal-affinity chromatography was used to obtain a highly purified detergent-solubilized TVP fraction with a yield around 1.5 mg of protein per litre of yeast culture. Electron microscopy showed the monodispersity of the purified protein and single particle analysis provided the first direct evidence of a dimeric structure for H+-PPases. We propose that the method developed could be useful for large-scale purification of other recombinant thermophilic membrane proteins.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Biomembranes - Volume 1716, Issue 1, 1 October 2005, Pages 69-76
Journal: Biochimica et Biophysica Acta (BBA) - Biomembranes - Volume 1716, Issue 1, 1 October 2005, Pages 69-76
نویسندگان
Rosa L. López-Marqués, José R. Pérez-Castiñeira, Morten J. Buch-Pedersen, Sergio Marco, Jean-Louis Rigaud, Michael G. Palmgren, Aurelio Serrano,