Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Jun 5

PMID 24897378

Citations 4

Authors

Su-Lim Choi

Sang Jun Lee

Soo-Jin Yeom

Hyun Ju Kim

Young Ha Rhee

Heung-Chae Jung

Seung-Goo Lee

Affiliations

Soon will be listed here.

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 KDs of 8-1,000 µM 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.

Citing Articles

Exploring the use of leucine zippers for the generation of a new class of inclusion bodies for pharma and biotechnological applications.

Roca-Pinilla R, Fortuna S, Natalello A, Sanchez-Chardi A, Ami D, Aris A Microb Cell Fact. 2020; 19(1):175.

PMID: 32887587 PMC: 7650227. DOI: 10.1186/s12934-020-01425-x.

A new generation of recombinant polypeptides combines multiple protein domains for effective antimicrobial activity.

Roca-Pinilla R, Lopez-Cano A, Saubi C, Garcia-Fruitos E, Aris A Microb Cell Fact. 2020; 19(1):122.

PMID: 32503648 PMC: 7275485. DOI: 10.1186/s12934-020-01380-7.

Controlled Aggregation and Increased Stability of β-Glucuronidase by Cellulose Binding Domain Fusion.

Yeom S, Han G, Kim M, Kwon K, Fu Y, Kim H PLoS One. 2017; 12(1):e0170398.

PMID: 28099480 PMC: 5242468. DOI: 10.1371/journal.pone.0170398.

Engineering structure and function using thermoresponsive biopolymers.

Pastuszka M, MacKay J Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2015; 8(1):123-38.

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