Monoclonal antibody to paired helical filaments. Hiroshi MORI 1 1Department of Clinical Physiology, Tokyo Metropolitan Institute of Gerontology pp.339-345
Published Date 1987/4/10
DOI https://doi.org/10.11477/mf.1431905889
  • Abstract
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 Paired helical filaments (PHF), a distinct type of filaments, are the principal constituent of neurofibrillary tangles and neurites in senile plaques in the cerebral cortex of the subjects with Alzheimer's disease. The insolubility of PHF in various solvents enabled us to obtain highly purified PHF, but prevented the conventional analysis of their components. Here I reported that a hybridoma producing antibody (DF2) against PHF has been obtained after fusion between mouse myeloma and spleen cells from a Lewis rat immunized with purified PHF. DF2 has been found to recognize PHF and 5 kDa protein by the immunoblot study, suggesting that 5 kDa protein is a component of PHF. On the basis of immunoreactivity of DF2, antigenic 5 kDa protein was purified from human brain with molecular-sieving chromatography and HPLC. Purified 5 kDa protein was sequenced and proved to be ubiquitin. Vice versa, affinity-purified antibody to ubiquitin is found to decorate PHF at light and electromicroscopic level. Furthermore, ubiquitin sequence was directly identified in the proteolytic digestion of PHF. The epitope of DF2 was also determined by means of dot-immunobinding test using synthetic peptides and proteolytic fragments derived from ubiquitin. This epitope was found to occur near the /3 structure in the hydrophilic region by computer-aided hydrophobicity profile analysis and the secondary structure prediction.

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