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

doi:10.1016/j.jsb.2012.11.006

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

Molecular Pathobiology

Research output: Contribution to journal › Article › peer-review

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 language	English (US)
Pages (from-to)	190-194
Number of pages	5
Journal	Journal of Structural Biology
Volume	181
Issue number	2
DOIs	https://doi.org/10.1016/j.jsb.2012.11.006
State	Published - Feb 2013

Keywords

Affinity grid
Cryo-EM
PKC βII
RACK1
Ribosome

ASJC Scopus subject areas

Structural Biology

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10.1016/j.jsb.2012.11.006

Cite this

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title = "Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome",

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.",

keywords = "Affinity grid, Cryo-EM, PKC βII, RACK1, Ribosome",

author = "Gyanesh Sharma and Jesper Pallesen and Sanchaita Das and Robert Grassucci and Robert Langlois and Hampton, {Cheri M.} and Kelly, {Deborah F.} and {des Georges}, Amedee and Joachim Frank",

year = "2013",

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doi = "10.1016/j.jsb.2012.11.006",

language = "English (US)",

volume = "181",

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journal = "Journal of Structural Biology",

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T1 - Affinity grid-based cryo-EM of PKC binding to RACK1 on the ribosome

AU - Sharma, Gyanesh

AU - Pallesen, Jesper

AU - Das, Sanchaita

AU - Grassucci, Robert

AU - Langlois, Robert

AU - Hampton, Cheri M.

AU - Kelly, Deborah F.

AU - des Georges, Amedee

AU - Frank, Joachim

PY - 2013/2

Y1 - 2013/2

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AB - 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.

KW - Affinity grid

KW - Cryo-EM

KW - PKC βII

KW - RACK1

KW - Ribosome

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

Abstract

Keywords

ASJC Scopus subject areas

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