ALG-2 Attenuates COPII Budding In Vitro and Stabilizes the Sec23/Sec31A Complex
Figure 4
Sec31A is recruited to liposomes and Sec23 in an ALG-2-dependent manner.
A. Recombinant proteins bound to artificial liposomes and collected from a sucrose gradient were analyzed for recruitment efficiency in the presence of ALG-2 and a peptide representing the ABD (pABD). Antibodies directed against Sec31A, HA tag (HA-Sec13), Flag-tag (flag-Sec23A), Sar1 and ALG-2 were used to analyze protein recruitment. The immunoblot is representative of more than three independent experiments. B. Quantification of band intensities relative to Ca2+ control (ctr) in representative immunoblot. C. Anti-Flag antibody coated beads were used as bait for flag-Sec23. Protein binding was performed in the presence of Ca2+ and proteins in gels were visualized using SyproRed staining.