The hydrophobic amino acid residues of a denatured protein molecule tend to react with the particles of the stationary phase of hydrophobic interaction chromatography (STHIC). These hydrophobic interactions prevent th...The hydrophobic amino acid residues of a denatured protein molecule tend to react with the particles of the stationary phase of hydrophobic interaction chromatography (STHIC). These hydrophobic interactions prevent the denatured protein molecules from aggregating with each other. The STHIC can provide high enough energy to a denatured protein molecule to make it dehydration and to refold it into its native or various intermediate states. The outcome not only depends on the specific interactions between amino acids, the structure of STHIC, but also depends on the association between the STHIC and mobile phase. The mechanism of protein refolding and the principle of its quality control by HPHIC were also presented. By appropriate selection of the Chromatographie condition, several denatured proteins can be refolded and separated simultaneously in a single Chromatographic run. A specially designed unit, with diameter much larger than its length, was designed and employed for both laboratory and preparative scales. That unit for the simultaneous renaturation and purification of proteins (USRPP) had the following four functions: to completely remove denaturant, to renature proteins, to separate renatured proteins from impurities, and to easily recycle waste denaturant. The efficiencies of refolding and purification of proteins by the USRPP are almost comparable to a usual long Chromatographie column in laboratory. In preparative scale, USRPP can be easily, rapidly, and economically applied requiring a low pressure gradient. As an example, recombinant human interferon-γ is employed to elucidate the application of the preparative USRPP.展开更多
To date,few studies have reported on the folding mechanism of tandem G-quadruplexes in human telomeric DNA.Hence,the control of the biofunctions of G-quadruplex,which requires a thorough understanding of its dynamic b...To date,few studies have reported on the folding mechanism of tandem G-quadruplexes in human telomeric DNA.Hence,the control of the biofunctions of G-quadruplex,which requires a thorough understanding of its dynamic behavior,is limited.Here,we investigated the folding/unfolding behavior of human telomeric sequences with lengths over 10 kilonucleotide(knt)by circular dichroism(CD)spectroscopy,UV melting assay,and atomic force microscopy(AFM)-based single-molecule force spectroscopy.展开更多
基金the National Natural Science Foundation of China，the National Basic Research Program of China，the Foundation for Scholar of Hefei Normal University，the Foundations of Educational Committee of Anhui Province
基金This work was supposed by the National Natural ScienceFoundation of China (Grant Nos.39880003 and 20175016).
文摘The hydrophobic amino acid residues of a denatured protein molecule tend to react with the particles of the stationary phase of hydrophobic interaction chromatography (STHIC). These hydrophobic interactions prevent the denatured protein molecules from aggregating with each other. The STHIC can provide high enough energy to a denatured protein molecule to make it dehydration and to refold it into its native or various intermediate states. The outcome not only depends on the specific interactions between amino acids, the structure of STHIC, but also depends on the association between the STHIC and mobile phase. The mechanism of protein refolding and the principle of its quality control by HPHIC were also presented. By appropriate selection of the Chromatographie condition, several denatured proteins can be refolded and separated simultaneously in a single Chromatographic run. A specially designed unit, with diameter much larger than its length, was designed and employed for both laboratory and preparative scales. That unit for the simultaneous renaturation and purification of proteins (USRPP) had the following four functions: to completely remove denaturant, to renature proteins, to separate renatured proteins from impurities, and to easily recycle waste denaturant. The efficiencies of refolding and purification of proteins by the USRPP are almost comparable to a usual long Chromatographie column in laboratory. In preparative scale, USRPP can be easily, rapidly, and economically applied requiring a low pressure gradient. As an example, recombinant human interferon-γ is employed to elucidate the application of the preparative USRPP.
基金funded by the National Natural Science Foundation of China(nos.21827805 and 21525418).
文摘To date,few studies have reported on the folding mechanism of tandem G-quadruplexes in human telomeric DNA.Hence,the control of the biofunctions of G-quadruplex,which requires a thorough understanding of its dynamic behavior,is limited.Here,we investigated the folding/unfolding behavior of human telomeric sequences with lengths over 10 kilonucleotide(knt)by circular dichroism(CD)spectroscopy,UV melting assay,and atomic force microscopy(AFM)-based single-molecule force spectroscopy.
