Protein refolding assisted by self-assembled nanogels as novel artificial molecular chaperone.
Molecular chaperone-like activity for protein refolding was investigated using nanogels of self-assembly of cholesterol-bearing pullulan. Nanogels effectively prevented protein aggregation (i.e. carbonic anhydrase and citrate synthase) during protein refolding from GdmCl denaturation. Enzyme activity recovered in high yields upon dissociation of the gel structure in which the proteins were trapped, by the addition of cyclodextrins. The nanogels assisted protein refolding in a manner similar to the mechanism of molecular chaperones, namely by catching and releasing proteins. The nanogels acted as a host for the trapping of refolded intermediate proteins. Cyclodextrin is an effector molecule that controls the binding ability of these host nanogels to proteins. The present nanogel system was also effective at the renaturation of inclusion body of a recombinant protein of the serine protease family.
Nomura
Yuta
Y
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida-Hommachi, Sakyo-ku, 606-8501 Kyoto, Japan.
Ikeda
Masahiro
M
Yamaguchi
Nozomi
N
Aoyama
Yasuhiro
Y
Akiyoshi
Kazunari
K
D000818:Animals; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D002950:Citrate (si)-Synthase / Q000737:chemistry* / Q000378:metabolism; D003505:Cyclodextrins / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics; D005936:Glucans / Q000737:chemistry; D019791:Guanidine / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry*; D002479:Inclusion Bodies / Q000737:chemistry / Q000235:genetics; D007700:Kinetics; D018832:Molecular Chaperones / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013552:Swine
D003505:Cyclodextrins; D029968:Escherichia coli Proteins; D005936:Glucans; D020100:Hydrogels; D018832:Molecular Chaperones; D011994:Recombinant Proteins; C009109:pullulan; D002950:Citrate (si)-Synthase; D002256:Carbonic Anhydrases; D019791:Guanidine