Detergent Insoluble Proteins and Inclusion Body-Like Structures Immunoreactive for PRKDC/DNA-PK/DNA-PKcs, FTL, NNT, and AIFM1 in the Amygdala of Cognitively Impaired Elderly Persons

• Published in: Journal of neuropathology and experimental neurology Vol. 77; no. 1; pp. 21 - 39
• Main Authors: Gal, Jozsef, Chen, Jing, Katsumata, Yuriko, Fardo, David W, Wang, Wang-Xia, Artiushin, Sergey, Price, Douglas, Anderson, Sonya, Patel, Ela, Zhu, Haining, Nelson, Peter T
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.01.2018; by American Association of Neuropathologists, Inc
• Subjects: Aged; Aged, 80 and over; Alzheimer Disease - metabolism; Alzheimer Disease - pathology; Amygdala - metabolism; Amygdala - pathology; Apoferritins - metabolism; Apoptosis Inducing Factor - metabolism; Chromatography, Liquid; Cognitive Dysfunction - metabolism; Cognitive Dysfunction - pathology; Deoxyribonucleic acid; DNA; DNA-Activated Protein Kinase - metabolism; DNA-Binding Proteins - metabolism; Humans; Inclusion Bodies - metabolism; Inclusion Bodies - pathology; Ions; Mitochondrial Proteins - metabolism; Molecular weight; NADP Transhydrogenase, AB-Specific - metabolism; Nuclear Proteins - metabolism; Original; Proteins; Proteomics; Tandem Mass Spectrometry; LC-MS; Phosphorylation; GWAS; Proteomics; Tauopathy; ADSP; Lewy
• ISSN: 0022-3069; 1554-6578; 1554-6578
• DOI: 10.1093/jnen/nlx097

Abstract Misfolded protein in the amygdala is a neuropathologic feature of Alzheimer disease and many other neurodegenerative disorders. We examined extracts from human amygdala (snap-frozen at autopsy) to investigate whether novel and as yet uncharacterized misfolded proteins would be detectable. Polypeptides from the detergent-insoluble, urea-soluble protein fractions of amygdala were interrogated using liquid chromatography-electrospray ionization-tandem mass spectrometry. Among the detergent-insoluble proteins identified in amygdala of demented subjects but not controls were Tau, TDP-43, Aβ, α-synuclein, and ApoE. Additional detergent-insoluble proteins from demented subjects in the high-molecular weight portion of SDS gels included NNT, TNIK, PRKDC (DNA-PK, or DNA-PKcs), ferritin light chain (FTL), AIFM1, SYT11, STX1B, EAA1, COL25A1, M4K4, CLH1, SQSTM, SYNJ1, C3, and C4. In follow-up immunohistochemical experiments, NNT, TNIK, PRKDC, AIFM1, and FTL were observed in inclusion body-like structures in cognitively impaired subjects' amygdalae. Double-label immunofluorescence revealed that FTL and phospho-PRKDC immunoreactivity colocalized partially with TDP-43 and/or Tau inclusion bodies. Western blots showed high-molecular weight "smears", particularly for NNT and PRKDC. A preliminary genetic association study indicated that rare NNT, TNIK, and PRKDC gene variants had nominally significant association with Alzheimer-type dementia risk. In summary, novel detergent-insoluble proteins, with evidence of proteinaceous deposits, were found in amygdalae of elderly, cognitively impaired subjects.