Structural analysis of protein inclusion bodies by Fourier transform infrared microspectroscopy.
The expression of recombinant human growth hormone (h-GH) and human interferon-alpha-2b (IFN-alpha-2b) in E. coli leads to the formation of insoluble protein aggregates or inclusion bodies (IBs). The secondary structure of these IBs, their corresponding native forms and thermal aggregates were studied by Fourier Transform Infrared (FT-IR) spectroscopy and microspectroscopy. It was demonstrated that residual native-like structures were maintained within IBs at different extents depending on the level of expression, with possible implications in biotechnology. Furthermore, comparison between infrared spectra of thermal aggregates and IBs suggests new insights on the structure of protein aggregates.
Ami
Diletta
D
Dipartimento di Biotecnologie e Bioscienze, Università di Milano Bicocca, Piazza della, Scienza 2, 20126 Milan, Italy.
Natalello
Antonino
A
Taylor
Geoffrey
G
Tonon
Giancarlo
G
Maria Doglia
Silvia
S
D004926:Escherichia coli / Q000378:metabolism; D006358:Hot Temperature; D019382:Human Growth Hormone / Q000096:biosynthesis; D006801:Humans; D002479:Inclusion Bodies / Q000648:ultrastructure*; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D019382:Human Growth Hormone