دورية أكاديمية
Use of sequential chemical extractions to purify nuclear membrane proteins for proteomics identification.
| العنوان: | Use of sequential chemical extractions to purify nuclear membrane proteins for proteomics identification. |
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| المؤلفون: | Korfali N; The Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, UK., Fairley EA, Swanson SK, Florens L, Schirmer EC |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2009; Vol. 528, pp. 201-25. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Validation Study |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Print ISSN: 1064-3745 (Print) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | Cell Fractionation/*methods , Membrane Proteins/*isolation & purification , Nuclear Envelope/*chemistry , Proteomics/*methods, Alkalies/chemistry ; Chemical Fractionation/methods ; Chromatography, High Pressure Liquid ; Detergents/chemistry ; Humans ; Lymphocytes/chemistry ; Lymphocytes/cytology ; Microsomes/chemistry ; Salts/chemistry ; Software ; Tandem Mass Spectrometry |
| مستخلص: | The nuclear envelope (NE) is a double membrane system that is both a part of the endoplasmic reticulum and part of the nucleus. As its constituent proteins tend to be highly complexed with nuclear and cytoplasmic components, it is notoriously difficult to purify. Two methods can reduce this difficulty for the identification of nuclear membrane proteins: comparison to contaminating membranes and chemical extractions to enrich for certain groups of proteins. The purification of nuclear envelopes and contaminating microsomal membranes is described here along with procedures for chemical extraction using salt and detergent, chaotropes, or alkaline solutions. Each extraction method enriches for different combinations of nuclear envelope proteins. Finally, we describe the analysis of these fractions with MudPIT, a proteomics methodology that avoids gel extraction of bands to facilitate identification of minor proteins and membrane proteins that do not resolve well on gels. Together these three approaches can significantly increase the output of proteomics studies aimed at identifying the protein complement of subcellular membrane systems. |
| معلومات مُعتمدة: | 076616 United Kingdom WT_ Wellcome Trust |
| المشرفين على المادة: | 0 (Alkalies) 0 (Detergents) 0 (Membrane Proteins) 0 (Salts) |
| تواريخ الأحداث: | Date Created: 20090121 Date Completed: 20090319 Latest Revision: 20211020 |
| رمز التحديث: | 20240829 |
| DOI: | 10.1007/978-1-60327-310-7_15 |
| PMID: | 19153695 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1064-3745 |
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| DOI: | 10.1007/978-1-60327-310-7_15 |