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Cell-free Generation of COPII-coated Procollagen I Carriers
Lin Yuan1, Satoshi Baba1, Kanika Bajaj1
1Department of Molecular and Cell Biology and Howard Hughes Medical Institute, University of California, Berkeley, CA, USA.
Abstract:
The aim of this protocol is to generate COPII-coated procollagen I (PC1) carriers in a cell-free reaction. The COPII-coated PC1 carriers were reconstituted from donor membrane, cytosol, purified recombinant COPII proteins, and nucleotides. This protocol describes the preparation of donor membrane and cytosol, the assembly of the reaction, and the isolation and detection of reconstituted COPII-coated carriers. This cell-free reaction can be used to test conditions that stimulate or suppress the packaging of PC1 into COPII-coated carriers.
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