tailieunhanh - Báo cáo khoa học: Concepts and tools to exploit the potential of bacterial inclusion bodies in protein science and biotechnology

Cells have evolved complex and overlapping mechanisms to protect their proteins from aggregation. However, several reasons can cause the failure of such defences, among them mutations, stress conditions and high rates of protein synthesis, all common consequences of heterologous protein pro-duction. | IFEBS Journal MINIREVIEW Concepts and tools to exploit the potential of bacterial inclusion bodies in protein science and biotechnology Pietro Gatti-Lafranconi1 Antonino Natalello2 z Diletta Ami2 Silvia Maria Doglia2 and Marina Lotti2 1 Department of Biochemistry University of Cambridge UK 2 Department of Biotechnology and Biosciences State University of Milano-Bicocca Italy Keywords aggregation amyloid-like structures biocatalysis electron and optical microscopies fourier transform infrared spectroscopy inclusion bodies IB structural properties native-like conformation recombinant proteins stress response Correspondence S. M. Doglia M. Lotti Department of Biotechnology and Biosciences State University of Milano-Bicocca Piazza della Scienza 2 20126 Milano Italy Fax 39 02 64483565 Tel 39 02 64483459 E-mail Cells have evolved complex and overlapping mechanisms to protect their proteins from aggregation. However several reasons can cause the failure of such defences among them mutations stress conditions and high rates of protein synthesis all common consequences of heterologous protein production. As a result in the bacterial cytoplasm several recombinant proteins aggregate as insoluble inclusion bodies. The recent discovery that aggregated proteins can retain native-like conformation and biological activity has opened the way for a dramatic change in the means by which intracellular aggregation is approached and exploited. This paper summarizes recent studies towards the direct use of inclusion bodies in biotechnology and for the detection of bottlenecks in the folding pathways of specific proteins. We also review the major biophysical methods available for revealing fine structural details of aggregated proteins and which information can be obtained through these techniques. These authors contributed equally to this work Received 28 January 2011 revised 20 March 2011 accepted 5 April 2011 doi .