NSF, Unc-18-1, dynamin-1 and HSP90 are inclusion body components in neuronal intranuclear inclusion disease identified by anti-SUMO-1-immunocapture.
Neuronal intranuclear inclusion disease, a progressive ataxia that may be familial or sporadic, is characterized by numerous neuronal intranuclear inclusion bodies similar to those found in polyglutamine repeat diseases. Previously, we found that the intranuclear inclusion bodies are intensely immunopositive for SUMO-1, a protein which covalently conjugates to other proteins in a similar way to ubiquitin. To identify the SUMO-1-associated proteins in the inclusion bodies, we isolated intranuclear inclusion bodies from fresh, frozen brain tissue of a case with familial neuronal intranuclear inclusion disease and solubilized the proteins. SUMO-1-associated inclusion body proteins were then immunocaptured using an anti-SUMO-1 antibody. The proteins, NSF, dynamin-1 and Unc-18-1 (rbSEC1), involved in membrane trafficking of proteins, and the chaperone HSP90, were identified following anti-SUMO-1-immunocapture by using tandem mass spectrometry and database searching. Immunohistochemistry of brain sections and crude brain homogenates of three cases of familial neuronal intranuclear inclusion disease confirmed the presence of these proteins in intranuclear inclusions.
School of Medical Science, Griffith Institute of Health and Medical Research, Griffith University, Gold Coast Campus, Queensland, QLD, 4222, Australia. d.pountney@griffith.edu.au
Dean L
DL
Pountney
Mark J
MJ
Raftery
Fariba
F
Chegini
Peter C
PC
Blumbergs
Wei Ping
WP
Gai
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002467:Cell Nucleus / Q000378:metabolism / Q000473:pathology; D034284:Dynamin I / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D008854:Microscopy, Electron; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D019636:Neurodegenerative Diseases / Q000378:metabolism* / Q000473:pathology; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D025842:SUMO-1 Protein / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
C079091:APBA1 protein, human; D048868:Adaptor Proteins, Signal Transducing; D015415:Biomarkers; D018841:HSP90 Heat-Shock Proteins; D009419:Nerve Tissue Proteins; D025842:SUMO-1 Protein; D034284:Dynamin I; D052066:N-Ethylmaleimide-Sensitive Proteins