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, Deb Kelly, Amedee des Georges, Joachim Frank

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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
Volume181
Issue number2
DOIs
StatePublished - Feb 1 2013

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Ribosomes
Small Ribosome Subunits
Macromolecular Substances
Proteins

All Science Journal Classification (ASJC) codes

  • Structural Biology

Cite this

Sharma, G., Pallesen, J., Das, S., Grassucci, R., Langlois, R., Hampton, C. M., ... Frank, J. (2013). Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome. Journal of Structural Biology, 181(2), 190-194. https://doi.org/10.1016/j.jsb.2012.11.006
Sharma, Gyanesh ; Pallesen, Jesper ; Das, Sanchaita ; Grassucci, Robert ; Langlois, Robert ; Hampton, Cheri M. ; Kelly, Deb ; des Georges, Amedee ; Frank, Joachim. / Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome. In: Journal of Structural Biology. 2013 ; Vol. 181, No. 2. pp. 190-194.
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Sharma, G, Pallesen, J, Das, S, Grassucci, R, Langlois, R, Hampton, CM, Kelly, D, des Georges, A & Frank, J 2013, 'Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome', Journal of Structural Biology, vol. 181, no. 2, pp. 190-194. https://doi.org/10.1016/j.jsb.2012.11.006

Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome. / Sharma, Gyanesh; Pallesen, Jesper; Das, Sanchaita; Grassucci, Robert; Langlois, Robert; Hampton, Cheri M.; Kelly, Deb; des Georges, Amedee; Frank, Joachim.

In: Journal of Structural Biology, Vol. 181, No. 2, 01.02.2013, p. 190-194.

Research output: Contribution to journalArticle

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Sharma G, Pallesen J, Das S, Grassucci R, Langlois R, Hampton CM et al. Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome. Journal of Structural Biology. 2013 Feb 1;181(2):190-194. https://doi.org/10.1016/j.jsb.2012.11.006