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Co-immunoprecipitation Assay Using Endogenous Nuclear Proteins from Cells Cultured Under Hypoxic Conditions
Published on: August 2, 2018
Nogo-A couples with Apg-1 through interaction and co-ordinate expression under hypoxic and oxidative stress
Florian Kern1, Ruslan I Stanika, Bettina Sarg
1*Biocenter, Division of Neurobiochemistry, Innsbruck Medical University, 6020 Innsbruck, Austria.
The Biochemical Journal
|August 6, 2013
Summary
Nogo-A, a nerve growth inhibitor, interacts with heat-shock protein Apg-1. This protein interaction is crucial for neuronal responses to hypoxic and oxidative stress, impacting conditions like stroke and neuroblastoma.
Area of Science:
- Neuroscience
- Molecular Biology
- Cellular Stress Response
Background:
- Nogo-A (reticulon 4) is a key inhibitor of nerve regeneration in the central nervous system (CNS).
- Nogo-A is highly expressed in principal CNS neurons, with its specificity linked to the NiG domain.
Purpose of the Study:
- To identify novel interactors of the Nogo-A NiG domain.
- To investigate the role of Nogo-A in neuronal response to hypoxic and oxidative stress.
Main Methods:
- Affinity precipitation, co-immunoprecipitation, and proximity ligation assays were used.
- Experiments utilized primary hippocampal neurons from Nogo-deficient mice.
- Protein expression was analyzed under hypoxic and oxidative stress conditions.
Main Results:
- Apg-1, a heat-shock protein 110 kDa family member, was identified as a novel, selective interactor of Nogo-A's NiG domain.
- Nogo-A and Apg-1 showed tight co-regulation in hippocampal neurons under hypoxic and oxidative stress.
- Both proteins were upregulated during hypoxia but downregulated with hydrogen peroxide exposure.
Conclusions:
- Nogo-A is involved in the neuronal response to hypoxic and oxidative stress.
- The Nogo-A-Apg-1 interaction may be relevant for understanding stroke-induced ischemia and neuroblastoma formation.
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