Abstract
The conversion of prion protein (PrP) from the monomeric cellular isoform to the oligomeric pathological isoform is a crucial event in the pathogenesis of prion diseases. To investigate oligomer formation of PrP, enhanced green fluorescent protein (EGFP)-tagged PrP (EGFP-PrP) without the glycosylphosphatidylinositol (GPI) anchor was prepared and the oligomerization of EGFP-PrP induced by sodium dodecyl sulphate (SDS) was monitored by fluorescence correlation spectroscopy (FCS). The FCS analysis indicated that soluble oligomers were formed at 0.011% SDS. Furthermore, the combination of fluorescence cross-correlation spectroscopy (FCCS) and a panel of anti-PrP monoclonal antibodies (mAbs) revealed the conformational changes in PrP. Our studies provide a method to analyze conformational changes of proteins in solution.
Keywords: Fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy, prion protein, soluble oligomer, antibody, conformational change, epitope mapping
Current Pharmaceutical Biotechnology
Title: Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy
Volume: 11 Issue: 1
Author(s): Hiroshi Sakata, Motohiro Horiuchi, Izumi Takahashi and Masataka Kinjo
Affiliation:
Keywords: Fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy, prion protein, soluble oligomer, antibody, conformational change, epitope mapping
Abstract: The conversion of prion protein (PrP) from the monomeric cellular isoform to the oligomeric pathological isoform is a crucial event in the pathogenesis of prion diseases. To investigate oligomer formation of PrP, enhanced green fluorescent protein (EGFP)-tagged PrP (EGFP-PrP) without the glycosylphosphatidylinositol (GPI) anchor was prepared and the oligomerization of EGFP-PrP induced by sodium dodecyl sulphate (SDS) was monitored by fluorescence correlation spectroscopy (FCS). The FCS analysis indicated that soluble oligomers were formed at 0.011% SDS. Furthermore, the combination of fluorescence cross-correlation spectroscopy (FCCS) and a panel of anti-PrP monoclonal antibodies (mAbs) revealed the conformational changes in PrP. Our studies provide a method to analyze conformational changes of proteins in solution.
Export Options
About this article
Cite this article as:
Sakata Hiroshi, Horiuchi Motohiro, Takahashi Izumi and Kinjo Masataka, Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy, Current Pharmaceutical Biotechnology 2010; 11 (1) . https://dx.doi.org/10.2174/138920110790725357
DOI https://dx.doi.org/10.2174/138920110790725357 |
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
-
Ginkgo Biloba Extract (EGb 761) in Alzheimers Disease: Is there Any Evidence?
Current Alzheimer Research In Vitro and In Vivo Models of Multiple Sclerosis
CNS & Neurological Disorders - Drug Targets Targeting SKCa Channels in Cancer: Potential New Therapeutic Approaches
Current Medicinal Chemistry Cancer Prevention with Promising Natural Products: Mechanisms of Action and Molecular Targets
Anti-Cancer Agents in Medicinal Chemistry Reposition of the Fungicide Ciclopirox for Cancer Treatment
Recent Patents on Anti-Cancer Drug Discovery Sera/Organ Lysates of Selected Animals Living in Polluted Environments Exhibit Cytotoxicity against Cancer Cell Lines
Anti-Cancer Agents in Medicinal Chemistry Nature Against Depression
Current Medicinal Chemistry Bioinformatic Application in Proteomic Research on Biomarker Discovery and Drug Target Validation
Current Bioinformatics Targeting ErbB Receptors in High-Grade Glioma
Current Pharmaceutical Design Molecular Link Mechanisms between Inflammation and Cancer
Current Pharmaceutical Design Caspase-Independent Pathways of Programmed Cell Death: The Unraveling of New Targets of Cancer Therapy?
Current Cancer Drug Targets Phytochemical-Mediated Glioma Targeted Treatment: Drug Resistance and Novel Delivery Systems
Current Medicinal Chemistry Gene Expression Analysis Approach to Establish Possible Links Between Parkinson's Disease, Cancer and Cardiovascular Diseases
CNS & Neurological Disorders - Drug Targets Sulfur Containing Acridine Derivatives in Preclinical Studies with Cancer Cell Lines
Current Medicinal Chemistry Focus on the Multimodal Role of Biomarkers in Breast Cancer
Current Pharmaceutical Design New Improved Drug Delivery Technologies for Pentacyclic Triterpenes: A Review
Protein & Peptide Letters Immune-Inflammatory Responses and Oxidative Stress in Alzheimers Disease: Therapeutic Implications
Current Pharmaceutical Design Clostridial Neurotoxins: Mode of Substrate Recognition and Novel Therapy Development
Current Protein & Peptide Science Applications of 211At and 223Ra in Targeted Alpha-Particle Radiotherapy
Current Radiopharmaceuticals Pleiotrophin as a Possible New Target for Angiogenesis-Related Diseases and Cancer
Recent Patents on Anti-Cancer Drug Discovery