title: Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies.
abstract: Recombinant protein overexpression of large proteins in bacteria often results in insoluble and misfolded proteins directed to inclusion bodies. We report the application of shear stress in micrometer-wide, thin fluid films to refold boiled hen egg white lysozyme, recombinant hen egg white lysozyme, and recombinant caveolin-1. Furthermore, the approach allowed refolding of a much larger protein, cAMP-dependent protein kinase A (PKA). The reported methods require only minutes, which is more than 100 times faster than conventional overnight dialysis. This rapid refolding technique could significantly shorten times, lower costs, and reduce waste streams associated with protein expression for a wide range of industrial and research applications.
authors.affiliation: University of California, Irvine, Department of Molecular Biology and Biochemistry, Irvine, CA, 92697-2025 (USA).
authors.forename: Tom Z
authors.initials: TZ
authors.lastname: Yuan
authors.forename: Callum F G
authors.initials: CF
authors.lastname: Ormonde
authors.forename: Stephan T
authors.initials: ST
authors.lastname: Kudlacek
authors.forename: Sameeran
authors.initials: S
authors.lastname: Kunche
authors.forename: Joshua N
authors.initials: JN
authors.lastname: Smith
authors.forename: William A
authors.initials: WA
authors.lastname: Brown
authors.forename: Kaitlin M
authors.initials: KM
authors.lastname: Pugliese
authors.forename: Tivoli J
authors.initials: TJ
authors.lastname: Olsen
authors.forename: Mariam
authors.initials: M
authors.lastname: Iftikhar
authors.forename: Colin L
authors.initials: CL
authors.lastname: Raston
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Weiss
mesh_terms: D020134:Catalytic Domain; D051242:Caveolin 1 / Q000737:chemistry / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D004867:Equipment Design; D055772:Green Chemistry Technology / Q000295:instrumentation; D002479:Inclusion Bodies / Q000378:metabolism*; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D058849:Protein Refolding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D051242:Caveolin 1; D011994:Recombinant Proteins; D017868:Cyclic AMP-Dependent Protein Kinases; C421864:hen egg lysozyme; D009113:Muramidase
keywords: green chemistry; protein expression; protein folding; shear stress