title: The role of detergent in refolding of GdnHCl-denatured arginine kinase from shrimp Fenneropenaeus Chinensis: the solubilization of aggregate and refolding in detergent solutions.
abstract: Strong aggregation occurred in the refolding route of arginine kinase (AK) denatured with 3 mol GdnHCl/L (GdnHCl, guanidine hydrochloride). The activity recovery of GdnHCl-denatured AK was very low and dependent on the protein concentration in the process of refolding. For denatured AK at 1.2 micromol/L concentration, the recovered activity yield was about 45.2% of the native enzyme, whereas at 5.2 micromol/L the activity recovery yield was only 20% of native activity. The nonionic detergent Triton X-100 and Tween 20 (< or = 100 mmol/L concentration) not only effectively blocked the aggregation but also enabled the denatured AK to recover most of its native activity. The kinetics of aggregate solubilization showed that there was an induction phase dependent on the detergent, but there was no dependency when detergent was absent. The apparent activity recovery had a cooperative relation with detergents in the process of refolding, which suggested the existence of some interaction between the detergent and the refolding intermediate. On the basis of the study results, a scheme of refolding was proposed.
authors.affiliation: Department of Biology, Hubei Normal University, Huangshi, PR China.
authors.forename: Ji-Cheng
authors.initials: JC
authors.lastname: Pan
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Wang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Cheng
authors.forename: Zhenhang
authors.initials: Z
authors.lastname: Yu
authors.forename: Xue-Ming
authors.initials: XM
authors.lastname: Rao
authors.forename: Hai-Meng
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001122:Arginine Kinase / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D033364:Decapoda / Q000201:enzymology*; D003902:Detergents / Q000378:metabolism*; D019791:Guanidine / Q000494:pharmacology*; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D012995:Solubility
chemical_list: D003902:Detergents; D001122:Arginine Kinase; D019791:Guanidine