Subcellular analysis of aberrant protein structure in age-related neurodegenerative disorders.
Subcellular interactions of neurodegenerative disease proteins may provide a basic molecular mechanism underlying neuronal disorders. Each protein may also exhibit activities related to normal cell structure and function. It may be necessary to develop methodologies to distinguish between normal and abnormal intracellular interactions of such proteins in human cells. The latter would result in distinct perturbations in cell function depending both on the specific protein or nucleic acid interactions as well as its subcellular localization. Individual neurodegenerative disorders may be characterized by distinct alterations in subcellular neuronal protein structure and function. We developed as a basic experimental paradigm a novel human cell system to identify and examine such abnormal neuronal protein structures. The basic rationale is that neurodegenerative protein interactions would result in the formation of intracellular high molecular weight (HMW) complexes in cells from afflicted individuals. Following cell fractionation these unique structures could be detected by gradient sedimentation coupled with immunoblot analysis. They would not be observed in age matched control normal human cells. We now report that this procedure has been successfully used to determine a unique subcellular alteration of beta-amyloid precursor protein (beta-APP) structure in Alzheimer's disease (AD) cells. The latter was not observed in normal cells. Similar structural alterations were observed for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a protein known to bind to beta-APP in vitro. The utility of this model system to interrelate aberrant protein interactions of neurodegenerative disease proteins and their subcellular localization is considered.
Mazzola
Jennifer L
JL
Scientific Connexions, 105 Terry Drive, Suite 118, Newtown, PA 18940, USA.
Sirover
Michael A
MA
D000375:Aging / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000737:chemistry / Q000378:metabolism*; D001921:Brain / Q000166:cytology; D016022:Case-Control Studies; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D019636:Neurodegenerative Diseases / Q000378:metabolism*; D011487:Protein Conformation; D013347:Subcellular Fractions / Q000378:metabolism
D016564:Amyloid beta-Protein Precursor; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases