A method for rapidly screening functionality of actin mutants and tagged actins
العنوان: | A method for rapidly screening functionality of actin mutants and tagged actins |
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المؤلفون: | Heidi Rommelaere, Katrien Neirynck, Joël Vandekerckhove, Christophe Ampe, Davy Waterschoot |
المصدر: | Biological Procedures Online, Vol 6, Iss 1, Pp 235-249 (2004) Biological Procedures |
بيانات النشر: | Springer Science and Business Media LLC, 2004. |
سنة النشر: | 2004 |
مصطلحات موضوعية: | Genetics, lcsh:R5-920, biology, Biochemistry, Genetics and Molecular Biology(all), Binding sites, Mutant, macromolecular substances, Transfection, Actins, General Biochemistry, Genetics and Molecular Biology, Cell biology, lcsh:Biology (General), Chaperone (protein), biology.protein, Native state, Protein folding, Actin-binding protein, lcsh:Medicine (General), Cytoskeleton, lcsh:QH301-705.5, Actin, Research Article |
الوصف: | Recombinant production and biochemical analysis of actin mutants has been hampered by the fact that actin has an absolute requirement for the eukaryotic chaperone CCT to reach its native state. We therefore have developed a method to rapidly screen the folding capacity and functionality of actin variants, by combining in vitro expression of labelled actin with analysis on native gels, band shift assays or copolymerization tests. Additionally, we monitor, using immuno-fluorescence, incorporation of actin variants in cytoskeletal structures in transfected cells. We illustrate the method by two examples. In one we show that tagged versions of actin do not always behave native-like and in the other we study some of the molecular defects of three &bgr;-actin mutants that have been associated with diseases. |
تدمد: | 1480-9222 |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::49988b1a58efb48b0156d1104df76f5b https://doi.org/10.1251/bpo94 |
حقوق: | OPEN |
رقم الأكسشن: | edsair.doi.dedup.....49988b1a58efb48b0156d1104df76f5b |
قاعدة البيانات: | OpenAIRE |
تدمد: | 14809222 |
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