Escherichia coli small heat shock proteins IbpA-B enhance activity of enzymes sequestered in inclusion bodies

• Autorzy: Kuczynska-Wisnik D. , Zurawa-Janicka D. , Narkiewicz J. , Kwiatkowska J. , Lipinska B. , Laskowska E.
• Języki publikacji: EN

Abstrakty

EN

Escherichia coli small heat shock proteins, IbpA/B, function as molecular chape- rones and protect misfolded proteins against irreversible aggregation. IbpA/B are in­duced during overproduction of recombinant proteins and bind to inclusion bodies in E. coli cells. We investigated the effect of AibpA/B mutation on formation of inclu­sion bodies and biological activity of enzymes sequestered in the aggregates in E. coli cells. Using three different recombinant proteins: Cro-ß-galactosidase, ß-lactamase and rat rHtrA1 we demonstrated that deletion of the ibpA/B operon did not affect the level of produced inclusion bodies. However, in aggregates containing IbpA/B a higher enzymatic activity was detected than in the IbpA/B-deficient inclusion bodies. These results confirm that IbpA/B protect misfolded proteins from inactivation in vivo.

Słowa kluczowe

EN

IbpA protein; inclusion body; heat shock; IbpB protein; enzyme activity; Escherichia coli; protein aggregation

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2004
• Tom: 51
• Numer: 4
• Opis fizyczny: p.925-931,fig.,ref.

Twórcy

autor

Kuczynska-Wisnik D.

• University of Gdansk, Kladki 24, 80-822 Gdansk, Poland

autor

Zurawa-Janicka D.

autor

Narkiewicz J.

autor

Kwiatkowska J.

autor

Lipinska B.

autor

Laskowska E.

Bibliografia

• Allen SP, Polazzi JO, Gierse JK, Easton AM. (1992) Two novel heat shock genes encoding proteins produced in response to heterologous protein expression in E. coli. JBacteriol.; 174: 6938-47.
• Baneyx F. (1999) Recombinant protein expression in E. coli. Curr Opin Biotechnol.; 10: 411-21.
• Bowden GA, Paredes AM, Georgiou G. (1991) Structure and morfology of protein inclusion bodies in E. coli. Biotechnology..; 9: 725-30.
• Bradford MM. (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem.; 72: 248-54.
• Bressan GM, Stanley KK. (1987) pUEX, a bacterial expression vector related to pEX with universal host specifity. Nucleic Acids Res.; 15: 10056. .
• Carrio MM, Villaverde A. (2002) Construction and deconstruction of bacterial inclusion bodies. J Biotechnol.; 96: 3-12.
• Ehrnsperger M, Graber S, Gaestel M, Buchner J. (1997) Binding of non-native protein to hsp25 during heat shock creates a reservoir of folding intermediates for reactivation. EMBO J.; 16: 221-9.
• Hoffmann F, Rinas U. (2000) Kinetics of heat-shock response and inclusion body formation during temperature-induced production of basic fibroblast growth factor in high-cell-density cultures of recombinant E. coli. Biotechnol Prog.; 16: 1000-7.
• Jurgen B, Lin HY, Riemschneider S, Scharf C, Neubauer P, Schmid R, Hecker M, Schweder T. (2000) Monitoring of genes that respond to overproduction of an insoluble recombinant protein in E. coli glucose- limited fed-batch fermentation. BiotechnolBioeng; 70: 217-24.
• Kitagawa M, Miyakawa M, Matsumura Y, Tsuchido T. (2002) E. coli small heat shock proteins, IbpA and IbpB, protect enzymes from inactivation by heat and oxidants. Eur J Biochem.; 269: 2907-17.
• Kuczynska-Wisnik D, Kedzierska S, Matuszewska E, Lund P, Taylor A, Lipinska B, Laskowska E. (2002) The Escherichia coli small heat-shock proteins IbpA and IbpB prevent the aggregation of endogenous proteins denatured in vivo during extreme heat shock. Microbiology.; 148: 1757-65.
• Laskowska E, Wawrzynow A, Taylor A. (1996) IbpA and IbpB, the new heat-shock proteins, bind to endogenous Escherichia coli proteins aggregated intracellularly by heat shock. Biochimie.; 78: 117-22.
• Laskowska E, Kuczynska-Wisnik D, Bak M, Lipinska B. (2003) Trimethoprim induces heat shock proteins and protein aggregation in E. coli cells. Curr Microbiol.; 47: 286-9.
• Lee GJ, Vierling E. (2000) A small heat shock protein cooperates with heat shock protein 70 system to reactivate a heat denatured protein. Plant Physiol.; 122: 189-98.
• Meerman HJ, Georgiou G. (1994) Construction and characterization of a set of E. coli strains deficient in all known loci affecting the proteolytic stability of secreted recombinant proteins. Bio Technology.; 12: 1107-10.
• Mogk A, Deurling E, Vorderfulbecke S, Vierling E, Bukau B. (2003 a) Small heat shock proteins, ClpB and the DnaK system form a functional triade in reversing protein aggregation. MolMicrobiol.; 50: 585-95.
• Mogk A, Schlieker C, Friedrich KL, Schonfeld HJ, Vierling E, Bukau B. (2003b) Refolding of substrates bound to small Hsps relies on a diaggregation reaction mediated most efficiently by ClpB/DnaK. J Biol Chem.; 278: 31033-42.
• Narberhaus F. (2000) alpha-Crystallin-type heat shock proteins: socializing minichaperones in the context of a multichaperone network. Microbiol Mol Biol Rev.; 66: 64-93.
• Rinas U. (1996) Synthesis rates of cellular proteins involved in translation and protein folding are strongly altered in response to overproduction of basic fibroblast growth factor by recombinant E. coli. Biotechnol Prog.; 12: 196-200.
• Sambrook J, Fritsh EF, Maniatis F. (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
• Veinger L, Diamant S, Buchner J, Goloubinoff P. (1998) The small heat-shock protein IbpB from E. coli stabilizes stress-denatured proteins for subsequent refolding by a multichaperone network. J Biol Chem.; 273: 11032-7.