Tandem Detergent-Extraction and Immunoprecipitation of Proteinopathy (TDiP): Scalable Enrichment of ALS-Associated TDP-43 Aggregates
Year: 2022
Type: article
Abstract: Transactive response DNA-binding protein of 43 kDa (TDP-43) is a highly conserved, ubiquitously expressed nucleic acid-binding protein that regulates DNA/RNA metabolism. Genetics and neuropathology studies have linked TDP-43 to several neuromuscular and neurological disorders including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Under pathological conditions, TDP-43 mislocalizes to the cytoplasm where it forms insoluble, hyper-phosphorylated aggregates during disease progression. Here, we optimized a scalable in vitro immuno-purification strategy referred to as tandem detergent-extraction and immunoprecipitation of proteinopathy (TDiP) to isolate TDP-43 aggregates that recapitulate those identified in postmortem ALS tissue. Moreover, we demonstrate that these purified aggregates can be utilized in biochemical, proteomics, and live-cell assays. This platform offers a rapid, accessible, and streamlined approach to study ALS disease mechanisms, while overcoming many limitations that have hampered TDP-43 disease modeling and therapeutic drug discovery efforts.
Source: iScience
Institution: University of North Carolina at Chapel Hill
Language: English
Subfield: Clinical Neurology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $3,000
Funders: National Science Foundation, U.S. Department of Veterans Affairs, University of North Carolina, National Institutes of Health, Directorate for STEM Education, National Institute on Aging, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Neurological Disorders and Stroke, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill
Awards: G1623066935, Neural network dysfunction in early HIV neuropathogenesis, G2481947521, Dual CHIP Functions Control Tau Triage In Alzheimer's Disease, Dissecting the Role of HDAC6 in Alzheimer's Pathogenesis, G3941989759, Investigation of the innate and adaptive immune responses to TDP-43 aggregates in Amyotrophic Lateral Sclerosis, G4822179308, Graduate research Fellowship Program (GRFP), G5294160854, G529887834, G7366156696, G7989876034, Elucidating The Aberrant TDP-43 Species That Promote Neurodegeneration, A novel mouse model of TDP-43 Proteinopathy in FTLD-ALS: Elucidating the role of TDP-43 acetylation in neurodegeneration and proteostasis impairment, G8892363406, G9000033364, UNC-CH Summer Research Training in Aging for Medical Students, Composition and Function of Membrane Contact Sites in Differentiation and Disease, Training Program in Cellular Systems and Integrative Physiology, Cancer Center Support Grant, G958207224, G992476094
OpenAlex ID: W4364378042