| Products/Services Used | Details | Operation |
|---|---|---|
| Proteins, Expression, Isolation and Analysis | The Erf2–Erf4 complex was eluted with 5–6 column volumes of elution buffer (20 mM Tris–HCl pH 8.0, 150 mM NaCl, 0.02% DDM and 300 mM imidazole), and incubated with anti-FLAG tag affinity resin (Genscript) for 2 h at 4 °C. | Get A Quote |
Palmitoylation of cysteine residues at the C-terminal hypervariable regions in human HRAS and NRAS, which is necessary for RAS signaling, is catalyzed by the acyltransferase DHHC9 in complex with its accessory protein GCP16. The molecular basis for the acyltransferase activity and the regulation of DHHC9 by GCP16 is not clear. Here we report the cryo-electron microscopy structures of the human DHHC9-GCP16 complex and its yeast counterpart-the Erf2-Erf4 complex, demonstrating that GCP16 and Erf4 are not directly involved in the catalytic process but stabilize the architecture of DHHC9 and Erf2, respectively. We found that a phospholipid binding to an arginine-rich region of DHHC9 and palmitoylation on three resi... More
