کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10536757 | 962600 | 2016 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Yeast membrane proteomics using leucine metabolic labelling: Bioinformatic data processing and exemplary application to the ER-intramembrane protease Ypf1
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کلمات کلیدی
ERADLC-MSPSMEndoplasmic reticulum associated degradation - تداخل وابسته به اندوپلاسمی رتیکولومFeature detection - تشخیص ویژگیSILAC - سیلکendoplasmic reticulum - شبکه آندوپلاسمی liquid chromatography–mass spectrometry - طیف سنجی جرم کروماتوگرافی مایعIsotope labelling - نشانه گذاری ایزوتوپProteolysis - پروتئولیز
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
We describe in detail the usage of leucine metabolic labelling in yeast in order to monitor quantitative proteome alterations, e.g. upon removal of a protease. Since laboratory yeast strains are typically leucine auxotroph, metabolic labelling with trideuterated leucine (d3-leucine) is a straightforward, cost-effective, and ubiquitously applicable strategy for quantitative proteomic studies, similar to the widely used arginine/lysine metabolic labelling method for mammalian cells. We showcase the usage of advanced peptide quantification using the FeatureFinderMultiplex algorithm (part of the OpenMS software package) for robust and reliable quantification. Furthermore, we present an OpenMS bioinformatics data analysis workflow that combines accurate quantification with high proteome coverage. In order to enable visualization, peptide-mapping, and sharing of quantitative proteomic data, especially for membrane-spanning and cell-surface proteins, we further developed the web-application Proteator (http://proteator.appspot.com). Due to its simplicity and robustness, we expect metabolic leucine labelling in yeast to be of great interest to the research community. As an exemplary application, we show the identification of the copper transporter Ctr1 as a putative substrate of the ER-intramembrane protease Ypf1 by yeast membrane proteomics using d3-leucine isotopic labelling.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1864, Issue 10, October 2016, Pages 1363-1371
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1864, Issue 10, October 2016, Pages 1363-1371
نویسندگان
Lars Nilse, Dönem Avci, Patrick Heisterkamp, Oliver Serang, Marius K. Lemberg, Oliver Schilling,