Lon and ClpP proteases participate in the physiological disintegration of bacterial inclusion bodies.
Aggregated protein is solubilized by the combined activity of chaperones ClpB, DnaK and small heat-shock proteins, and this could account, at least partially, for the physiological disintegration of bacterial inclusion bodies. In vivo, the involvement of proteases in this process had been suspected but not investigated. By using an aggregation prone beta-galactosidase fusion protein produced in Escherichia coli, we show in this study that the main ATP-dependent proteases Lon and ClpP participate in the physiological disintegration of cytoplasmic inclusion bodies, their absence minimizing the protein removal up to 40%. However, the role of these proteases is clearly distinguishable especially regarding the fate of solubilized protein. While Lon appears as a minor contributor in the disintegration process, ClpP directs an important attack on the released or releasable protein even not being irreversibly misfolded. ClpP is then observed as a wide-spectrum, main processor of aggregation-prone proteins and also of polypeptides physiologically released from inclusion bodies, even when occurring as soluble versions with a conformation compatible with their enzymatic activity.
Vera
Andrea
A
Institut de Biotecnologia i de Biomedicina, Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain.
Arís
Anna
A
Carrió
Mar
M
González-Montalbán
Nuria
N
Villaverde
Antonio
A
D049071:Endopeptidase Clp / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology* / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D002479:Inclusion Bodies / Q000201:enzymology / Q000502:physiology*; D049070:Protease La / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012995:Solubility; D013347:Subcellular Fractions; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; C483567:Lon protein, E coli; D049070:Protease La; C483572:ClpP protease, E coli; D049071:Endopeptidase Clp