دورية أكاديمية
Detection of phosphorylated T and B cell antigen receptor species by Phos-tag SDS- and Blue Native-PAGE.
| العنوان: | Detection of phosphorylated T and B cell antigen receptor species by Phos-tag SDS- and Blue Native-PAGE. |
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| المؤلفون: | Deswal S; Department for Molecular Immunology, Max Planck-Institute for Immunobiology, Freiburg, Germany., Beck-García K, Blumenthal B, Dopfer EP, Schamel WW |
| المصدر: | Immunology letters [Immunol Lett] 2010 May 04; Vol. 130 (1-2), pp. 51-6. Date of Electronic Publication: 2009 Dec 22. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier/North-Holland Biomedical Press Country of Publication: Netherlands NLM ID: 7910006 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0542 (Electronic) Linking ISSN: 01652478 NLM ISO Abbreviation: Immunol Lett Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam, Elsevier/North-Holland Biomedical Press. |
| مواضيع طبية MeSH: | Multiprotein Complexes/*chemistry , Receptor-CD3 Complex, Antigen, T-Cell/*chemistry , Receptors, Antigen, B-Cell/*chemistry, Animals ; Blotting, Western ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Manganese/chemistry ; Mice ; Molecular Structure ; Phosphorylation ; Rosaniline Dyes/chemistry |
| مستخلص: | Detection of phospho-proteins and differently phosphorylated forms of the same protein are important in understanding cell behaviour. One novel method is Phos-tag SDS-PAGE. A dinuclear Mn(2+) complex that binds to phosphate groups (the Phos-tag) is covalently attached to the polyacrylamide gel matrix. Thus, phosphorylated proteins are retarded in their migration and can be distinguished from their non-phosphorylated counterparts. We applied Phos-tag SDS-PAGE to the analysis of the zeta, CD3epsilon and CD3delta subunits of the T cell antigen receptor (TCR-CD3). Pervanadate stimulation generated six different phospho-zeta and each two different CD3epsilon and CD3delta forms. This corresponds to the phosphorylatable tyrosines on their cytoplasmic tails. The phosphorylation pattern was compatible with random phosphorylation events. Further, we showed that the Phos-tag technology can be applied to Blue Native (BN)-PAGE. This extends the applicability to the analysis of native protein complexes. Upon pervanadate stimulation the TCR-CD3 complex was predominantly detected as two distinct phospho-complexes. In contrast, the B cell antigen receptor (BCR) appeared as one phospho-form. Thus, Phos-tag BN-PAGE is useful for the analysis of different phosphorylation states of multiprotein complexes. (Copyright 2009 Elsevier B.V. All rights reserved.) |
| المشرفين على المادة: | 0 (Multiprotein Complexes) 0 (Receptor-CD3 Complex, Antigen, T-Cell) 0 (Receptors, Antigen, B-Cell) 0 (Rosaniline Dyes) 42Z2K6ZL8P (Manganese) M1ZRX790SI (coomassie Brilliant Blue) |
| تواريخ الأحداث: | Date Created: 20091217 Date Completed: 20100827 Latest Revision: 20151119 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.imlet.2009.12.012 |
| PMID: | 20005898 |
| قاعدة البيانات: | MEDLINE |
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Full Text Finder | تدمد: | 1879-0542 |
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| DOI: | 10.1016/j.imlet.2009.12.012 |