Studies on Aggregation Interaction between Reduced Denatured Egg White Lysozymes during Refolding Procedure in Urea Solution

• Published in: Chinese journal of chemistry Vol. 25; no. 3; pp. 364 - 369
• Main Author: 边六交 梁长利 杨晓燕 刘莉
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.03.2007; WILEY‐VCH Verlag
• Subjects: aggregate; aggregation interaction; disulfide bond; non-covalent bond; reduced-denatured egg white lysozyme; 双硫键; 变性蛋白; 尿素溶液; 溶菌酶; 聚集作用; 重折叠
• ISSN: 1001-604X; 1614-7065
• DOI: 10.1002/cjoc.200790070

The aggregation interaction between reduced-denatured egg white lysozymes during refolding procedure in urea solution was studied by means of reducing and non-reducing protein electrophoreses. Results of non-reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis （SDS-PAGE） of the supernatant and aggregate precipitate formed in refolding process show that except being refolded to native egg white lysozymes, the reduced-denatured lysozymes can also form the aggregates with molecular weights （MW） being separately about 30.0 and 35.0 kD, while the reducing SDS-PAGE and the refolding results in the presence of sodium dodecyl sulphate show that these aggregates are formed chiefly through the misconnection of disulfide bonds between the reduced-denatured lysozymes, and the aggregate precipitates are formed through the non-covalent interactions between the aggregates with molecular weight being about 30.0 kD. From the results of electrophoresis and size-exclusion chromatographic analyses, it can be inferred that the aggregates with molecular weights being about 30.0 and 35.0 kD are bi-molecular and tri-molecular egg white lysozyme aggregates, respectively. And finally, a suggested refolding mechanism of reduced-denatured egg white lysozymes in urea solution was presented.