Folding aggregated proteins into functionally active forms.
The successful expression and purification of proteins in an active form is essential for structural and biochemical studies. With rapid advances in genome sequencing and high-throughput structural biology, an increasing number of proteins are being identified as potential drug targets but are difficult to obtain in a form suitable for structural or biochemical studies. Although prokaryotic recombinant expression systems are often used, proteins obtained in this way are typically found to be insoluble. Several experimental approaches have therefore been developed to refold these aggregated proteins into a biologically active form, often suitable for structural studies. The major refolding strategies adopt one of two approaches - chromatographic methods or refolding in free solution - and both routes have been successfully used to refold a range of proteins. Future advances are likely to involve the development of automated approaches for protein refolding and purification.
Swietnicki
Wieslaw
W
Integrated Toxicology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702, USA. wes.swietnicki@amedd.army.mil
D001709:Biotechnology / Q000379:methods; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D018832:Molecular Chaperones / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012997:Solvents / Q000737:chemistry
D018832:Molecular Chaperones; D011994:Recombinant Proteins; D012997:Solvents