Site-specific labeling of proteins for electron microscopy

Dambacher, Corey M.; Lander, Gabriel C.

doi:10.1016/j.jsb.2015.09.010

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Site-specific labeling of proteins for electron microscopy

Published Date:

Sep 25 2015

Publisher's site:

http://dx.doi.org/10.1016/j.jsb.2015.09.010 New Site Icon

Source:

J Struct Biol. 192(2):151-158.

[PDF - 1.03 MB]

Details:

Personal Authors:

Dambacher, Corey M. ; Lander, Gabriel C. ;

Keywords:

[+]

Pubmed ID:

26409249

Pubmed Central ID:

PMC4633364

Description:

Electron microscopy is commonly employed to determine the subunit organization of large macromolecular assemblies. However, the field lacks a robust molecular labeling methodology for unambiguous identification of constituent subunits. We present a strategy that exploits the unique properties of an unnatural amino acid in order to enable site-specific attachment of a single, readily identifiable protein label at any solvent-exposed position on the macromolecular surface. Using this method, we show clear labeling of a subunit within the 26S proteasome lid subcomplex that has not been amenable to labeling by traditional approaches.

Document Type:

Journal Article

Collection(s):

CDC Public Access

Funding:

DP2 EB020402/EB/NIBIB NIH HHS/United States
DP2 EB020402-01/DP/NCCDPHP CDC HHS/United States

Supporting Files:

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nihms728932.nxml	application/octet-stream
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nihms728932f2.jpg	image/jpeg

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