title: Controlled localization of functionally active proteins to inclusion bodies using leucine zippers.
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.
authors.affiliation: Biochemicals and Synthetic Biology Research Center, KRIBB, Yuseong-gu, Daejeon, Korea; Department of Bioscience and Biotechnology, Chungnam National University, Yuseong-gu, Daejeon, Korea.
authors.forename: Su-Lim
authors.initials: SL
authors.lastname: Choi
authors.affiliation: Infection and Immunity Biology Research Center, KRIBB, Yuseong-gu, Daejeon, Korea; Korea University of Science and Technology, Yuseong-gu, Daejeon, Korea.
authors.forename: Sang Jun
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Biochemicals and Synthetic Biology Research Center, KRIBB, Yuseong-gu, Daejeon, Korea.
authors.forename: Soo-Jin
authors.initials: SJ
authors.lastname: Yeom
authors.affiliation: Infection and Immunity Biology Research Center, KRIBB, Yuseong-gu, Daejeon, Korea; Korea University of Science and Technology, Yuseong-gu, Daejeon, Korea.
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Bioscience and Biotechnology, Chungnam National University, Yuseong-gu, Daejeon, Korea.
authors.forename: Young Ha
authors.initials: YH
authors.lastname: Rhee
authors.affiliation: Biochemicals and Synthetic Biology Research Center, KRIBB, Yuseong-gu, Daejeon, Korea.
authors.forename: Heung-Chae
authors.initials: HC
authors.lastname: Jung
authors.affiliation: Biochemicals and Synthetic Biology Research Center, KRIBB, Yuseong-gu, Daejeon, Korea; Korea University of Science and Technology, Yuseong-gu, Daejeon, Korea.
authors.forename: Seung-Goo
authors.initials: SG
authors.lastname: Lee
mesh_terms: D003600:Cytosol / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D002479:Inclusion Bodies / Q000378:metabolism*; D016350:Leucine Zippers / Q000502:physiology*; D011487:Protein Conformation