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

Jozsef Gal; Jing Chen; Yuriko Katsumata; David W. Fardo; Wang Xia Wang; Sergey Artiushin; Douglas Price; Sonya Anderson; Ela Patel; Haining Zhu; Peter T. Nelson

doi:10.1093/jnen/nlx097

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

Jozsef Gal, Jing Chen, Yuriko Katsumata, David W. Fardo, Wang Xia Wang, Sergey Artiushin, Douglas Price, Sonya Anderson, Ela Patel, Haining Zhu, Peter T. Nelson

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

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 detergentinsoluble, 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. Doublelabel 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.

Original language	English
Pages (from-to)	21-39
Number of pages	19
Journal	Journal of Neuropathology and Experimental Neurology
Volume	77
Issue number	1
DOIs	https://doi.org/10.1093/jnen/nlx097
State	Published - Jan 1 2018

Bibliographical note

Publisher Copyright:
© 2017 American Association of Neuropathologists, Inc.

Funding

Send correspondence to: Peter T. Nelson, MD, PhD, Division of Neuropa-thology, Department of Pathology, Rm 311, Sanders-Brown Center on Aging, University of Kentucky, 800 S. Limestone St., Lexington, KY 40536-0230; E-mail: [email protected] The study was supported by NIH grants P30 AG028383, R01 AG042419, R01 AG042475, T32 AG 000242, R21 NS095299 (to J.G.), and VA MERIT award I01 BX002149 (to H.Z.). The Orbitrap mass spectrometer was acquired by NIH Grant S10 RR029127 (to H.Z.).

Funders	Funder number
NIH NIDDK	P30 AG028383, T32 AG 000242, R01 AG042419, R21 NS095299, R01 AG042475
VA Merit Award	S10 RR029127, I01 BX002149
National Institute on Aging	P30AG028383
University of Kentucky

Keywords

ADSP
GWAS
LC-MS
Lewy
Phosphorylation
Proteomics
Tauopathy

ASJC Scopus subject areas

Pathology and Forensic Medicine
Neurology
Clinical Neurology
Cellular and Molecular Neuroscience

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1093/jnen/nlx097

Cite this

Gal, J., Chen, J., Katsumata, Y., Fardo, D. W., Wang, W. X., Artiushin, S., Price, D., Anderson, S., Patel, E., Zhu, H., & Nelson, P. T. (2018). 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. Journal of Neuropathology and Experimental Neurology, 77(1), 21-39. https://doi.org/10.1093/jnen/nlx097

@article{7820debecdc44b68ab502fb3fb227b40,

title = "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",

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 detergentinsoluble, 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. Doublelabel 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.",

keywords = "ADSP, GWAS, LC-MS, Lewy, Phosphorylation, Proteomics, Tauopathy",

author = "Jozsef Gal and Jing Chen and Yuriko Katsumata and Fardo, {David W.} and Wang, {Wang Xia} and Sergey Artiushin and Douglas Price and Sonya Anderson and Ela Patel and Haining Zhu and Nelson, {Peter T.}",

note = "Publisher Copyright: {\textcopyright} 2017 American Association of Neuropathologists, Inc.",

year = "2018",

month = jan,

day = "1",

doi = "10.1093/jnen/nlx097",

language = "English",

volume = "77",

pages = "21--39",

number = "1",

}

Gal, J, Chen, J, Katsumata, Y , Fardo, DW , Wang, WX, Artiushin, S, Price, D, Anderson, S, Patel, E, Zhu, H & Nelson, PT 2018, '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', Journal of Neuropathology and Experimental Neurology, vol. 77, no. 1, pp. 21-39. https://doi.org/10.1093/jnen/nlx097

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. / Gal, Jozsef; Chen, Jing; Katsumata, Yuriko et al.
In: Journal of Neuropathology and Experimental Neurology, Vol. 77, No. 1, 01.01.2018, p. 21-39.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - 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

AU - Gal, Jozsef

AU - Chen, Jing

AU - Katsumata, Yuriko

AU - Fardo, David W.

AU - Wang, Wang Xia

AU - Artiushin, Sergey

AU - Price, Douglas

AU - Anderson, Sonya

AU - Patel, Ela

AU - Zhu, Haining

AU - Nelson, Peter T.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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 detergentinsoluble, 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. Doublelabel 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.

AB - 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 detergentinsoluble, 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. Doublelabel 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.

KW - ADSP

KW - GWAS

KW - LC-MS

KW - Lewy

KW - Phosphorylation

KW - Proteomics

KW - Tauopathy

UR - http://www.scopus.com/inward/record.url?scp=85040176237&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85040176237&partnerID=8YFLogxK

U2 - 10.1093/jnen/nlx097

DO - 10.1093/jnen/nlx097

M3 - Article

C2 - 29186589

AN - SCOPUS:85040176237

SN - 0022-3069

VL - 77

SP - 21

EP - 39

JO - Journal of Neuropathology and Experimental Neurology

JF - Journal of Neuropathology and Experimental Neurology

IS - 1

ER -

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

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this