title: Increase of solubility of foreign proteins in Escherichia coli by coproduction of the bacterial thioredoxin.
abstract: Eukaryotic proteins are frequently produced in Escherichia coli as insoluble aggregates. This is one of the barriers to studies of macromolecular structure. We have examined the effect of coproduction of the E. coli thioredoxin (Trx) or E. coli chaperones GroESL on the solubility of various foreign proteins. The solubilities of all eight vertebrate proteins examined including transcription factors and kinases were increased dramatically by coproduction of Trx. Overproduction of E. coli chaperones GroESL increased the solubilities of four out of eight proteins examined. Although the tyrosine kinase Lck that was produced as an insoluble form and solubilized by urea treatment had a very low autophosphorylating activity, Lck produced in soluble form by coproduction of Trx had an efficient activity. These results suggest that the proteins produced in soluble form by coproduction of Trx have the native protein conformation. The mechanism by which coproduction of Trx increases the solubility of the foreign proteins is discussed.
authors.affiliation: Laboratory of Molecular Genetics, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Yasukawa
authors.forename: C
authors.initials: C
authors.lastname: Kanei-Ishii
authors.forename: T
authors.initials: T
authors.lastname: Maekawa
authors.forename: J
authors.initials: J
authors.lastname: Fujimoto
authors.forename: T
authors.initials: T
authors.lastname: Yamamoto
authors.forename: S
authors.initials: S
authors.lastname: Ishii
mesh_terms: D000818:Animals; D018835:Chaperonin 10 / Q000096:biosynthesis / Q000235:genetics; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000235:genetics; D018833:Chaperonins / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli; D005057:Eukaryotic Cells; D008967:Molecular Biology / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012995:Solubility; D013879:Thioredoxins / Q000096:biosynthesis* / Q000235:genetics; D014714:Vertebrates
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D011994:Recombinant Proteins; D013879:Thioredoxins; D018833:Chaperonins