| Artificial chaperone-assisted refolding of bovine carbonic anhydrase using molecular assemblies of stimuli-responsive polymers. | |
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MedLine Citation:
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PMID: 14606877 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An artificial chaperone, which can decrease the protein aggregation and increase the reactivation yield of denatured protein in a fashion similar to natural chaperone, was newly developed using stimuli-responsive polymers. It has previously been reported that the addition of poly(propylene oxide)-phenyl-poly(ethylene glycol) (PPOn-Ph-PEG) with the unit number of PPO (n) 33 could enhance the refolding of bovine carbonic anhydrase (Kuboi et al. J. Chromatogr. B 2000, 243, 213). PPO-Ph-PEG with a large PPO chain (n = 50) was synthesized and the surface properties were characterized by both the relative fluorescence intensity of 1-anilino-8-naphthalene sulfonate (ANS) and the fluidity determined by diphenylhexatriene (DPH). The variation of ANS intensity and DPH fluidity is shown in a diagram as functions of temperature and polymer concentration. The high values of ANS intensity and fluidity of PPO50-Ph-PEG were obtained in a relatively wide conditional range (more than 0.08 mM and more than 15 degrees C) although the conditions showing the high values of PPO33-Ph-PEG were restricted (more than 0.1 mM and more than 40 degrees C). It was also found that molecular assemblies of PPOn-Ph-PEG with diameters of 7-18 nm were formed in the above conditions. On the basis of the surface properties of their polymer self-assemblies, the possibility of using them as an artificial chaperone was investigated. The effect of the addition of PPOn-Ph-PEG on the reactivation yield of a model protein, carbonic anhydrase from bovine (CAB), and the optical density of the solution was examined at various temperatures and concentrations. The reactivation yield of CAB was strongly enhanced and the aggregate formation (the optical density) was suppressed by adding PPOn-Ph-PEG in the above conditions, which show high ANS intensity and DPH fluidity. Especially in the presence of 0.1 mM PPO50-Ph-PEG, the reactivation yield of CAB reached approximately 100% at 40-55 degrees C. It was thus found that self-assemblies of the present polymer could be utilized as an artificial chaperone by selecting suitable stimuli conditions. |
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Authors:
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Noriko Yoshimoto; Takuya Hashimoto; Matundu Menayame Felix; Hiroshi Umakoshi; Ryoichi Kuboi |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Biomacromolecules Volume: 4 ISSN: 1525-7797 ISO Abbreviation: Biomacromolecules Publication Date: 2003 Nov-Dec |
Date Detail:
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Created Date: 2003-11-10 Completed Date: 2004-07-26 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 100892849 Medline TA: Biomacromolecules Country: United States |
Other Details:
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Languages: eng Pagination: 1530-8 Citation Subset: IM |
Affiliation:
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Department of Chemical Science and Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka 560-8531, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Carbonic Anhydrases / chemistry*, drug effects Cattle Molecular Chaperones / pharmacology* Molecular Mimicry* Polyethylene Glycols / chemistry Polymers / chemistry*, pharmacology Propylene Glycols / chemistry Protein Engineering / methods Protein Renaturation / drug effects* Recombinant Proteins / chemistry, drug effects |
| Chemical | |
Reg. No./Substance:
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0/Molecular Chaperones; 0/Polyethylene Glycols; 0/Polymers; 0/Propylene Glycols; 0/Recombinant Proteins; 25322-69-4/polypropylene glycol; EC 4.2.1.1/Carbonic Anhydrases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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