title: Role of the disaggregase ClpB in processing of proteins aggregated as inclusion bodies.
abstract: Overproduction of heterologous proteins in bacterial systems often results in the formation of insoluble inclusion bodies (IBs), which is a major impediment in biochemical research and biotechnology. In principle, the activity of molecular chaperones could be employed to gain control over the IB formation and to improve the recombinant protein yields, but the potential of each of the major bacterial chaperones (DnaK/J, GroEL/ES, and ClpB) to process IBs has not been fully established yet. We investigated the formation of inclusion bodies (IBs) of two aggregation-prone proteins, VP1LAC and VP1GFP, overproduced in Escherichiacoli in the presence and absence of the chaperone ClpB. We found that both ClpB isoforms, ClpB95 and ClpB80 accumulated in E. coli cells during the production of IBs. The amount of IB proteins increased in the absence of ClpB. ClpB supported the resolubilization and reactivation of the aggregated VP1LAC and VP1GFP in E. coli cells. The IB disaggregation was optimal in the presence of both ClpB95 and ClpB80. Our results indicate an essential role of ClpB in controlling protein aggregation and inclusion body formation in bacteria.
authors.affiliation: Department of Biochemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Zblewska
authors.affiliation: Department of Biochemistry, University of Gdansk, 80-308 Gdansk, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Krajewska
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: Michal
authors.initials: M
authors.lastname: Zolkiewski
authors.affiliation: Department of Biochemistry, University of Gdansk, 80-308 Gdansk, Poland. Electronic address: sabina.kedzierska-mieszkowska@biol.ug.edu.pl.
authors.forename: Sabina
authors.initials: S
authors.lastname: Kędzierska-Mieszkowska
mesh_terms: D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D049071:Endopeptidase Clp; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D029544:Foot-and-Mouth Disease Virus; D005696:Galactosidases / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D036022:Capsid Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005696:Galactosidases; D049071:Endopeptidase Clp; C069775:ClpB protein, E coli
keywords: ClpB; Inclusion bodies; Molecular chaperone; Protein aggregation