Calpain-1 C2L domain peptide protects mouse hippocampus-derived neuronal HT22 cells against glutamate-induced oxytosis
| Autor: | Taku Ozaki, Takumi Abe, Ken Itoh, Mayu Sugawara, Shuya Kasai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
CPP
cell penetrating peptide Proteases QH301-705.5 Short Communication Biophysics PBS phosphate-buffered saline Mitochondrial calpain-1 Peptide QD415-436 Mitochondrion DMEM Dulbecco's Modified Eagle Medium Biochemistry Oxytosis Hippocampal HT22 cells medicine Neurodegeneration Biology (General) BBB blood–brain barrier Hn-μCL polyhistidine-conjugated μCL chemistry.chemical_classification Cell penetrating peptide biology MTS 3-(4 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium Glutamate receptor Calpain medicine.disease Cell biology AIF apoptosis-inducing factor ssDNA single stranded DNA chemistry Cytoplasm Cell-penetrating peptide biology.protein DAPI 4′ 6-diamidino-2-phenylindole |
| Zdroj: | Biochemistry and Biophysics Reports, Vol 27, Iss, Pp 101101-(2021) Biochemistry and Biophysics Reports |
| ISSN: | 2405-5808 |
| Popis: | Calpains are Ca2+-dependent cysteine proteases; their aberrant activation is associated with several neurodegenerative diseases. The μ-calpain catalytic subunit, calpain-1, is located in the cytoplasm as well as in the mitochondria. Mitochondrial calpain-1 cleaves apoptosis-inducing factor (AIF), leading to apoptotic cell death. We have previously reported that short peptides of calpain-1 C2-like domain conjugated with cell penetrating peptide HIV-Tat (Tat-μCL) selectively inhibit mitochondrial calpain-1 and effectively prevent neurodegenerative diseases of the eye. In this study, we determined whether mitochondrial calpain-1 mediates oxytosis (oxidative glutamate toxicity) in hippocampal HT22 cells using Tat-μCL and newly generated polyhistidine-conjugated μCL peptide and compared their efficacies in preventing oxytosis. TUNEL assay and single strand DNA staining revealed that both μCL peptides inhibited glutamate-induced oxytosis. Additionally, both the peptides suppressed the mitochondrial AIF translocation into the nucleus. All polyhistidine-μCL peptides (containing 4–16 histidine residues) showed higher cell permeability than Tat-μCL. Notably, tetrahistidine (H4)-μCL exerted the highest cytoprotective activity. Thus, H4-μCL may be a potential peptide drug for calpain-1-mediated neurodegenerative diseases such as Alzheimer's disease. Highlights • Mitochondrial calpain-1 mediates glutamate-induced oxytosis in HT22 cells. • CPP-μCL inhibits AIF translocation and DNA fragmentation in oxytosis. • Polyhistidine conjugation enhances intracellular μCL peptide uptake efficiency. |
| Databáze: | OpenAIRE |
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