Controlled localization of functionally active proteins to inclusion bodies using leucine zippers = 류신지퍼 이용 활성단백질 함유 단백질 봉입체 제조

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Title
Controlled localization of functionally active proteins to inclusion bodies using leucine zippers = 류신지퍼 이용 활성단백질 함유 단백질 봉입체 제조
Author(s)
Su-Lim Choi; Sang Jun Lee; Soo Jin Yeom; Hyun Ju Kim; Y H Rhee; Heung Chae JungSeung Goo Lee
Bibliographic Citation
PLoS One, vol. 9, no. 6, pp. e97093-e97093
Publication Year
2014
Abstract
Inclusion bodies (IBs) are typically non-functional particles of aggregated proteins. However, some proteins in fusion with amyloid-like peptides, viral coat proteins, and cellulose binding domains (CBDs) generate IB particles retaining the original functions in cells. Here, we attempted to generate CBD IBs displaying functional leucine zipper proteins (LZs) as bait for localizing cytosolic proteins in E. coli. When a red fluorescent protein was tested as a target protein, microscopic observations showed that the IBs red-fluoresced strongly. When different LZ pairs with KD s of 8-1,000 mM were tested as the bait and prey, the localization of the red fluorescence appeared to change following the affinities between the LZs, as observed by fluorescence imaging and flow cytometry. This result proposed that LZ-tagged CBD IBs can be applied as an in vivo matrix to entrap cytosolic proteins in E. coli while maintaining their original activities. In addition, easy detection of localization to IBs provides a unique platform for the engineering and analyses of protein-protein interactions in E. coli.
ISSN
1932-6203
Publisher
Public Library of Science
DOI
http://dx.doi.org/10.1371/journal.pone.0097093
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
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