Detection of prion protein oligomers by single molecule fluorescence imaging

Satoko Shibano, Kensuke Sasaki, Satoru Kidoaki, Toru Iwaki

Research output: Contribution to journalArticle

Abstract

The degree of polymerization of PrP has a close relationship with the pathological mechanisms of prion diseases. We examined, at the molecular level, the polymerization state of PrP in lysates of prion-infected cells using total internal reflection fluorescence microscopy (TIRFM). The crude lysates were fractionated by gel-filtration spin columns according to their molecular size. Both the oligomer-rich and the monomer-rich fractions were probed with fluorescein-labeled anti-PrP antibodies (mAb SAF70 and mAb 8G8). Fluorescent spots of varying intensity were detected, with the ratio of intense fluorescent spots being greater in the oligomer fraction samples with mAb SAF70 than those with 8G8, the specific epitope of which is thought to be buried in abnormal PrP molecules. The results indicated that PrP oligomers could be specifically detected and conformational changes of abnormal PrP molecules observed. Imaging by TIRFM may aid in determining the polymerization state and properties of PrP oligomers in pathological processes.

Original languageEnglish
Pages (from-to)1-6
Number of pages6
JournalNeuropathology
Volume33
Issue number1
DOIs
Publication statusPublished - Feb 1 2013

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Optical Imaging
Polymerization
Fluorescence Microscopy
Prion Diseases
Prions
Pathologic Processes
Fluorescein
Gel Chromatography
Epitopes
Anti-Idiotypic Antibodies
Single Molecule Imaging
Prion Proteins

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Clinical Neurology

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Detection of prion protein oligomers by single molecule fluorescence imaging. / Shibano, Satoko; Sasaki, Kensuke; Kidoaki, Satoru; Iwaki, Toru.

In: Neuropathology, Vol. 33, No. 1, 01.02.2013, p. 1-6.

Research output: Contribution to journalArticle

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