Analysis of Golgi complex functions: in vitro reconstitution systems
In vitro reconstitution is prerequisite to investigate complex cellular functions at the molecular level. Reconstitution systems range from combining complete cellular cytosol with organelle-enriched membrane fractions to liposomal systems where all components are chemically defined and can be chose...
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| Main Authors: | , |
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| Format: | Article (Journal) Chapter/Article |
| Language: | English |
| Published: |
26 November 2013
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| In: |
Methods for analysis of Golgi complex function
Year: 2013, Volume: 118, Pages: 3-14 |
| DOI: | 10.1016/B978-0-12-417164-0.00001-X |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/B978-0-12-417164-0.00001-X
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| Author Notes: | Frank Adolf and Felix Wieland |
| Summary: | In vitro reconstitution is prerequisite to investigate complex cellular functions at the molecular level. Reconstitution systems range from combining complete cellular cytosol with organelle-enriched membrane fractions to liposomal systems where all components are chemically defined and can be chosen at will. Here, we describe the in vitro reconstitution of COPI-coated vesicles from semi-intact cells. Efficient vesicle formation is achieved by simple incubation of permeabilized cells with the minimal set of coat proteins Arf1 and coatomer, and guanosine trinucleotides. GTP hydrolysis or any mechanical manipulations are not required for efficient COPI vesicle release. |
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| Item Description: | Gesehen am 06.05.2021 |
| Physical Description: | Online Resource |
| ISBN: | 9780124171831 0124171834 0124171648 9780124171640 |
| DOI: | 10.1016/B978-0-12-417164-0.00001-X |