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Title 

Controlled localization of functionally active proteins to inclusion bodies using leucine zippers

 

류신지퍼 이용 활성단백질 함유 단백질 봉입체 제조

Authors 

Su-Lim ChoiSang Jun LeeSoo Jin YeomHyun Ju KimY H RheeHeung Chae JungSeung Goo Lee

Publisher 

Public Library of Science

Issue Date 

2014

Citation 

Plos One, vol. 9, no. 6, pp. e97093-e97093

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

Link 

http://dx.doi.org/10.1371/journal.pone.0097093

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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