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

Gyanesh Sharma, Jesper Pallesen, Sanchaita Das, Robert Grassucci, Robert Langlois, Cheri M. Hampton, Deborah F. Kelly, Amedee des Georges, Joachim Frank

Research output: Contribution to journalArticlepeer-review


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 βII binds to the small ribosomal subunit (40S). We reconstructed a cryo-EM map which shows that PKC βII interacts with RACK1, a seven-bladed β-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 βII/RACK1 interaction and its role in translation.

Original languageEnglish (US)
Pages (from-to)190-194
Number of pages5
JournalJournal of Structural Biology
Issue number2
StatePublished - Feb 2013


  • Affinity grid
  • Cryo-EM
  • PKC βII
  • RACK1
  • Ribosome

ASJC Scopus subject areas

  • Structural Biology


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