title: Proteolytic digestion of bacterial inclusion body proteins during dynamic transition between soluble and insoluble forms.
abstract: Inclusion bodies formed by two closely related hybrid proteins, namely VP1LAC and LACVP1, have been compared during their building in Escherichia coli. Features of these proteins are determinant of aggregation rates and protein composition of the bodies, generating insoluble particles with distinguishable volume evolution. Interestingly, in LACVP1 and less perceptibly in VP1LAC bodies, an important fraction of the aggregated polypeptide is lost at a given stage of body construction. Stable degradation intermediates of the more fragile LACVP1 are concomitantly found embedded in the bodies. When recombinant protein synthesis is arrested in growing cells, the amount of aggregated protein drops while the amount of soluble protein undergoes a sudden rise before proteolysis. This indicates an architectural plasticity during the in vivo building of the studied inclusion bodies by a dynamic transition between soluble and insoluble forms of the recombinant proteins involved. During this transition, protease-sensitive polypeptides can suffer an efficient proteolytic attack and the resulting fragments further aggregate as inclusion body components.
authors.affiliation: Institut de Biologia Fonamental and Departament de Genètica and Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain.
authors.forename: M M
authors.initials: MM
authors.lastname: Carrió
authors.forename: J L
authors.initials: JL
authors.lastname: Corchero
authors.forename: A
authors.initials: A
authors.lastname: Villaverde
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D015153:Blotting, Western; D004926:Escherichia coli; D002479:Inclusion Bodies / Q000737:chemistry; D010957:Plasmids; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis; D012995:Solubility; D013997:Time Factors; D001616:beta-Galactosidase / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D001616:beta-Galactosidase