C-terminal 15 kDa fragment of cytoskeletal actin is posttranslationallyN-myristoylated upon caspase-mediated cleavage and targeted to mitochondria
| Autor: | Kengo Nakano, Rumi Ishisaka, Nagisa Sakurai, Toshihiko Utsumi |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Recombinant Fusion Proteins
Molecular Sequence Data Biophysics Apoptosis Cleavage (embryo) Myristic Acid Caspase substrate Biochemistry Structural Biology Chlorocebus aethiops Genetics medicine Animals Humans Staurosporine Amino Acid Sequence Actin-binding protein Cytoskeleton Molecular Biology Gelsolin Actin Caspase Myristoylation Posttranslational N-myristoylation Protein N-myristoylation biology Cell Biology Molecular biology Fusion protein Actins Peptide Fragments Mitochondria Protein acylation Protein Transport Caspases COS Cells biology.protein Posttranslational modification Carrier Proteins Protein Processing Post-Translational BH3 Interacting Domain Death Agonist Protein Plasmids medicine.drug |
| Zdroj: | FEBS Letters. 539:37-44 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/s0014-5793(03)00180-7 |
| Popis: | To detect the posttranslational N-myristoylation of caspase substrates, the susceptibility of the newly exposed N-terminus of known caspase substrates to protein N-myristoylation was evaluated by in vivo metabolic labeling with [3H]myristic acid in transfected cells using a fusion protein in which the query sequence was fused to a model protein. As a result, it was found that the N-terminal nine residues of the newly exposed N-terminus of the caspase-cleavage product of cytoskeletal actin efficiently direct the protein N-myristoylation. Metabolic labeling of COS-1 cells transiently transfected with cDNA coding for full-length truncated actin (tActin) revealed the efficient incorporation of [3H]myristic acid into this molecule. When COS-1 cells transiently transfected with cDNA coding for full-length actin were treated with staurosporine, an apoptosis-inducing agent, an N-myristoylated tActin was generated. Immunofluorescence staining coupled with MitoTracker or fluorescence tagged-phalloidin staining revealed that exogenously expressed tActin colocalized with mitochondria without affecting cellular and actin morphology. Taken together, these results demonstrate that the C-terminal 15 kDa fragment of cytoskeletal actin is posttranslationally N-myristoylated upon caspase-mediated cleavage during apoptosis and targeted to mitochondria. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text