Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome

作者：Sharma Gyanesh; Pallesen Jesper; Das Sanchaita; Grassucci Robert; Langlois Robert; Hampton Cheri M; Kelly Deborah F; des Georges Amedee; Frank Joachim^*

来源：Journal of Structural Biology, 2013, 181(2): 190-194.

DOI：10.1016/j.jsb.2012.11.006

• Affinity grid
• Cryo-EM
• RACK1
• PKC beta II
• Ribosome

Affinity grids (AG) are specialized EM grids that bind macromolecular complexes containing tagged proteins to obtain maximum occupancy for structural analysis through single-particle EM. In this study, utilizing AG, we show that His-tagged activated PKC beta II binds to the small ribosomal subunit (40S). We reconstructed a cryo-EM map which shows that PKC beta II interacts with RACK1, a seven-bladed beta-propeller protein present on the 40S and binds in two different regions close to blades 3 and 4 of RACK1. This study is a first step in understanding the molecular framework of PKC beta II/RACK1 interaction and its role in translation.

• 出版日期2013-2
• 单位Virginia Tech; 美国弗吉尼亚理工大学(Virginia Tech)