Protein quality in bacterial inclusion bodies

Protein aggregation in bacteria: the thin boundary between functionality and toxicity

“…The specific pole localization allows rapid elimination of the aggregated proteins from most of the cell population. Presence of IBs is directly associated with ageing and a decrease of cell productivity (Ventura & Villaverde, 2006). Therefore, IB formation might be a sophisticated mechanism to reduce the potentially toxic, partially folded monomers or small oligomers, of which the solvent-exposed APRs may interact with cellular components leading to exhaustion of the proteostatic machinery.…”

Inclusion body anatomy and functioning of chaperone-mediated in vivo inclusion body disassembly during high-level recombinant protein production in Escherichia coli

“…They have long been regarded as a localized mass of unordered polypeptide chains but more recent research suggested that these proteins may also have defined conformations within the aggregated state (Ventura and Villaverde, 2006). Inclusion bodies also have long 5 been considered inert towards in vivo dissolution, but it is now generally accepted that inclusion body proteins can be solubilized in vivo concomitant to or followed by proteolytic degradation (Corchero et al, 1997, Carrió et al, 1999Cubarsí et al, 2001;LeThanh et al, 2005;Vera et al, 2005) or even by folding into the native state LeThanh et al, 2005).…”

“…Goloubinoff et al, 1989;Blum et al, 1992;Georgiou and Valax, 1996;Nishihara et al, 1998;Baneyx and Palumbo, 2002). Although the substrate specificity of these chaperone systems 10 has been characterized and an enormous progress in understanding chaperone mediated protein folding processes has been achieved, the success of chaperone coproduction on increasing soluble protein accumulation is still a trial-and-error process and cannot be predicted from properties of the recombinant protein (Hoffmann and Rinas, 2004;Baneyx and Mujacic, 2004;Ventura and Villaverde, 2006). Moreover, in some cases, e.g.…”

Soluble Expression of Recombinant Human Smp30 for Detecting Serum Smp30 Antibody Levels in Hepatocellular Carcinoma Patients

“…Co-expression of chaperones with recombinant proteins has been traditionally used to increase the amount of soluble protein (Goloubinoff et al, 1989;Ventura et al, 2006). Because successful chaperone co-expression to improve solubility is a trial-and-error process that can not be predicted from properties of proteins (Baneyx et al, 2004;Hoffmann et al, 2004), we explored nine combinations of chaperones four chaperones with pET30a-SMP30 and five chaperones with pColdIII-SMP30, and found that SMP30 was rescued from aggregation by co -expression of chaperones pKJE7, pG-Tf2 and pTf16, especially pTf16.…”