Abstract
It has been a conventional notion that cytoplasmic recombinant expression leads to either soluble protein or inclusion bodies. In the latter case, it was always assumed that proteins in inclusion bodies (IBs) are more or less unfolded and hence require complete denaturing condition for solubilization, which uses strong detergents, urea or guanidine hydrochloride. However, we often observe distribution of expressed proteins in both soluble and insoluble fractions. In such expression, IBs are often loose and of flocculate morphology. We believe that such distribution is due to association of near native structures of the expressed proteins, which cause either aggregation into insoluble fractions or unstable soluble proteins. In our experience, although not reported by others, interleukin-1α, interferon-γ, tumor necrosis factors, fibroblast growth factors, His-tagged fyn kinase and many other proteins showed such behavior. If this occurs, we have experienced problems of instability, low yield and insolubility whether purification is done from the soluble fraction or by refolding of IBs. Arginine has shown great promise in non-denaturaing solubilization of some of these proteins we have tested.
Keywords: Non-denaturing solubilization, inclusion bodies, arginine, GFP, FGF-20
Current Pharmaceutical Biotechnology
Title: Non-Denaturing Solubilization of Inclusion Bodies
Volume: 11 Issue: 3
Author(s): Kouhei Tsumoto, Ryota Abe, Daisuke Ejima and Tsutomu Arakawa
Affiliation:
Keywords: Non-denaturing solubilization, inclusion bodies, arginine, GFP, FGF-20
Abstract: It has been a conventional notion that cytoplasmic recombinant expression leads to either soluble protein or inclusion bodies. In the latter case, it was always assumed that proteins in inclusion bodies (IBs) are more or less unfolded and hence require complete denaturing condition for solubilization, which uses strong detergents, urea or guanidine hydrochloride. However, we often observe distribution of expressed proteins in both soluble and insoluble fractions. In such expression, IBs are often loose and of flocculate morphology. We believe that such distribution is due to association of near native structures of the expressed proteins, which cause either aggregation into insoluble fractions or unstable soluble proteins. In our experience, although not reported by others, interleukin-1α, interferon-γ, tumor necrosis factors, fibroblast growth factors, His-tagged fyn kinase and many other proteins showed such behavior. If this occurs, we have experienced problems of instability, low yield and insolubility whether purification is done from the soluble fraction or by refolding of IBs. Arginine has shown great promise in non-denaturaing solubilization of some of these proteins we have tested.
Export Options
About this article
Cite this article as:
Tsumoto Kouhei, Abe Ryota, Ejima Daisuke and Arakawa Tsutomu, Non-Denaturing Solubilization of Inclusion Bodies, Current Pharmaceutical Biotechnology 2010; 11 (3) . https://dx.doi.org/10.2174/138920110791111924
DOI https://dx.doi.org/10.2174/138920110791111924 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
Call for Papers in Thematic Issues
Artificial Intelligence in Bioinformatics
Bioinformatics is an interdisciplinary field that analyzes and explores biological data. This field combines biology and information system. Artificial Intelligence (AI) has attracted great attention as it tries to replicate human intelligence. It has become common technology for analyzing and solving complex data and problems and encompasses sub-fields of machine ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Pyrrole: Chemical Synthesis, Microwave Assisted Synthesis, Reactions and Applications: A Review
Current Organic Chemistry Interleukin 20
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Bacteriophage Lambda as a Vehicle for Peptide and Protein Display
Current Pharmaceutical Biotechnology Daclatasvir and Sofosbuvir Mitigate Hepatic Fibrosis Through Downregulation of TNF-α / NF-κB Signaling Pathway
Current Molecular Pharmacology Third Generation Antipsychotic Drugs: Partial Agonism or Receptor Functional Selectivity?
Current Pharmaceutical Design G4 Aptamers: Trends in Structural Design
Mini-Reviews in Medicinal Chemistry Cadherins: The Superfamily Critically Involved in Breast Cancer
Current Pharmaceutical Design Pulmonary Vasodilators in the Management of Low Cardiac Output Syndrome After Pediatric Cardiac Surgery
Current Vascular Pharmacology Helminth Infections and Cardiovascular Diseases: Toxocara Species is Contributing to the Disease
Current Cardiology Reviews Artificial Carriers: A Strategy for Constructing Antigenic/Immunogenic Conjugates
Current Topics in Medicinal Chemistry MicroRNAs and Lung Cancer: New Oncogenes and Tumor Suppressors, New Prognostic Factors and Potential Therapeutic Targets
Current Medicinal Chemistry Future Targeted Disease Modifying Drugs for Alzheimer's Disease
Recent Patents on CNS Drug Discovery (Discontinued) Colostral Proline-Rich Polypeptides - Immunoregulatory Properties and Prospects of Therapeutic Use in Alzheimers Disease
Current Alzheimer Research Enhanced Oral Bioavailability of Oleanolic Acid in Rats with Phospholipid Complex
Letters in Drug Design & Discovery Bone Marrow Mesenchymal Stem Cells: Agents of Immunomodulation and Neuroprotection
Current Stem Cell Research & Therapy 3dswap-pred: Prediction of 3D Domain Swapping from Protein Sequence Using Random Forest Approach
Protein & Peptide Letters Editorial (Thematic Issue: Novel Insights on Aortic Aneurysm)
Current Pharmaceutical Design Patent Selections
Recent Patents on Biotechnology Aromatase: The Enzyme and Its Inhibition
Anti-Cancer Agents in Medicinal Chemistry Mechanisms Leading to Autoantibody Production: Link Between Inflammation and Autoimmunity
Current Drug Targets - Inflammation & Allergy