Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies.
Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies.
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DOI：
10.1002/cbic.201402427
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发表时间：
2015-02-09
期刊：
影响因子：
3.2
通讯作者：
Weiss, Gregory A.
中科院分区：
文献类型：
--
作者：
Yuan, Tom Z.;Ormonde, Callum F. G.;Kudlacek, Stephan T.;Kunche, Sameeran;Smith, Joshua N.;Brown, William A.;Pugliese, Kaitlin M.;Olsen, Tivoli J.;Iftikhar, Mariam;Raston, Colin L.;Weiss, Gregory A.
细菌中大蛋白质的重组蛋白过度表达通常会导致不溶性和错误折叠的蛋白质定向到包涵体。我们报道了在微米宽的薄液膜中应用剪切应力来重新折叠煮鸡蛋清溶菌酶、重组鸡蛋清溶菌酶和重组 Caveolin-1。此外，该方法允许更大的蛋白质、cAMP 依赖性蛋白激酶 A (PKA) 的重折叠。报道的方法只需要几分钟，比传统的过夜透析快 100 倍以上。这种快速重折叠技术可以显着缩短时间、降低成本并减少与蛋白质表达相关的废物流，适用于广泛的工业和研究应用。
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 the much larger protein, cAMP-dependent protein kinase A (PKA). The reported methods require only minutes, which is >100-times faster than conventional, overnight dialysis. This rapid refolding technique could significantly shorten the times, lower costs, and reduce the waste streams associated with protein expression for a wide-range of industrial and research applications.
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