[Studies on formation of aggregates from denatured lysozymes upon renaturing with size exclusion chromatography].
In order to study the renaturation mechanism of denatured protein in denaturant solution, the renaturation and separation process for three kinds of lysozyme molecules, which were separately denatured by urea and guanidine hydrochloride in the presence of reducing agents, was studied by size exclusion chromatography. When initial lysozyme concentration in denaturant solution was more than 10 g/L, the denatured lysozyme molecules were renatured and isolated in a size exclusion chromatographic column. A refolded lysozyme intermediate, a bi-molecular aggregate, was found. This result was confirmed by non-reducing sodium dodecyl sulfate-polyacrylamide gel electrohoresis (SDS-PAGE) analysis of renatured lysozyme molecules with the dilution method. Compared with the dilution method, the amount of the bi-molecular aggregate found by size exclusion chromatography was far less than that found in the dilution method. This result shows that the process for the renaturing of denatured lysozyme molecules in solution can be well described with three-state model in the presence of reducing agents.
Bian
Liujiao
L
Center of Gene-engineering, College of Life Science, Northwest University, Xi'an 710069, China. bianliujiao@sohu.com
Yang
Xiaoyan
X
Liu
Li
L
D000818:Animals; D002645:Chickens; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004527:Egg Proteins / Q000737:chemistry / Q000302:isolation & purification; D004529:Egg White / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D019791:Guanidine / Q000737:chemistry; D009113:Muramidase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*; D011489:Protein Denaturation; D017510:Protein Folding; D020673:Protein Renaturation; D014508:Urea / Q000737:chemistry
D004527:Egg Proteins; D014508:Urea; C421864:hen egg lysozyme; D009113:Muramidase; D019791:Guanidine