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Guanidine hydrochloride |50-01-1|—Essential reagents for protein Protein denaturation and renaturation

2017-03-09 来源:亚科官网
9 March 2017
  Guanidine hydrochloride can be used as an intermediate for pharmaceuticals, pesticides, dyes and other organic compounds, as well as a strong denaturant in RNA extraction of cells, and denaturation and renaturation of proteins. In this article, we will summarize the role of guanidine hydrochloride in protein denaturation and renaturation.
1. Guanidine hydrochloride can make protein denaturation
  Proteins are more susceptible to external factors (temperature, denaturants, etc.) and resulting in the conformational changes, loss of activity, abnormal changes of physical and chemical properties, this process is called protein denaturation. Guanidine hydrochloride is an important denaturant that produces a strong denaturing effect which includes two denatured mechanisms.
  One is the solubilization of guanidine hydrochloride on hydrophobic amino acid residues. Guanidine hydrochloride can break the hydrogen bonds in the protein structure, which increases the solubility of the nonpolar molecules, including the amino acid side chains, and reduces the hydrophobic interaction. Another mechanism is that the denatured proteins are preferentially combined with guanidine hydrochloride to form denatured proteins-denaturing agent complex, when the complex is removed, the reaction equilibrium moves to the right, and the protein in natural state is continuously converted into a complex that eventually leads to complete denaturation of the protein. However, the combination of denaturant and denatured protein is very weak, only high concentrations of denaturant can cause complete denaturation of the protein. In general, guanidine hydrochloride-induced denaturation is usually reversible.
2. Protein renaturation
  If the condition of degeneration is intense, the denaturation of the protein is irreversible. Some denatured proteins, when the denatured factor is removed, can be automatically restored to the natural state, this phenomenon is known as protein renaturation. In the process of protein renaturation, the inclusion body protein is first dissolved. Inclusion body protein refers to the protein expressed in the cell agglutinate and form amorphous, non-water-soluble, inactive solid particles, which is only soluble in denaturants such as urea, guanidine hydrochloride and so on.
  After the slow removal of the denaturant, the target protein is restored from the fully extended state of the denatured to the normal folded structure, and the reducing agent is removed to make the disulfide bond form normally. The renaturation process is generally start from the concentration of guanidine is 4M and end to 1.5M. There are several different ways:
  Renaturation by Dilution: add water or buffer directly into dilution solution, overnight, the protein can precipitate, the disadvantage is that the volume increased quickly, denaturant dilution rate is too fast, and not easy to control; dialysis or ultrafiltration: denaturant are small molecule material, in the dialysis or ultrafiltration process, they can be gradually penetrate through the porous membrane, and the concentration of denaturant in denatured protein solution gradually reduced, finally removed, so that the denatured protein can refold. The advantage is that the volume does not change, the speed can be controled, but the process is relatively slow; Column refolding: achieving refolding in the chromatographic process, the advantage is that the chromatographic fixation has a low adsorption capacity for the denatured protein, which can improve the refolding quality and the activity yield. At the same time, the target protein and the hybrid protein can be separated to achieve the purpose of purification, and the denaturant is easy to be collected.
  Proteins are life-related biological macromolecules, it is essential material to maintain the normal life and the second generation code for life genetics. A variety of research on protein is also a topic of interest, and its structural research is also important to promote medical progress. Protein denaturation or renaturation is one of the core problems in life science research. Among them, the denaturant guanidine hydrochloride not only can denature the protein, facilitate the study of its structure, but also can dissolve the inclusion body protein in the process of refolding, it plays an indispensable role in the study of life sciences.
Edited by the Editorial Office of Suzhou Yacoo Science Co., Ltd.