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

Biomedical Engineering

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

30 Scopus citations

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

All Science Journal Classification (ASJC) codes

Structural Biology

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10.1016/j.jsb.2012.11.006

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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.",

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",

note = "Funding Information: We thank Melissa Thomas for assistance with the illustrations. The work was supported by HHMI and grant R01 GM29169 (to JF). ",

year = "2013",

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

language = "English (US)",

volume = "181",

pages = "190--194",

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

N1 - Funding Information: We thank Melissa Thomas for assistance with the illustrations. The work was supported by HHMI and grant R01 GM29169 (to JF).

PY - 2013/2

Y1 - 2013/2

N2 - 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.

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.

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JO - Journal of Structural Biology

JF - Journal of Structural Biology

IS - 2

ER -

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

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