Aging and the Insolubleome: Identifying SDS-insoluble proteins from brains of aging and neurodegenerative disease mouse models by mass spectrometry.

摘要

Many age-related and neurological diseases show a dramatic increase in protein inclusion bodies, fibrils and/or aggregates in the brain. Even aged, but healthy brain tissue may exhibit insoluble protein inclusions compared to healthy young brain tissue, yet a comprehensive description of what constitutes these inclusions has not been carried out. Such insoluble protein aggregates by their very nature can impose technical difficulties in their analysis. We harvested brain material both from aged and young mice, as well as a Huntington's Disease (HD) mouse model (R6/2) that displays extensive nuclear insoluble aggregates compared to healthy controls. Recently, we have extended our (differential) analysis of insoluble protein aggregates to other species, including yeast strains (comparing young vs. aged and/or strains grown under caloric restriction vs. ad libitum) and also C. elegans (comparing long- or short-lived mutant strains to wild-type strains). In all cases, we employ a formic acid protocol [1] to solubilize SDS-insoluble protein fractions and then analyze the SDS-insoluble proteins ('insolubleome') using nano-HPLC-ESI-MS/MS (QSTAR) and vMALDI-MS/MS (vMALDI-LTQ) mass spectrometric technologies.