FEBS Letters | |
C‐terminal 15 kDa fragment of cytoskeletal actin is posttranslationally N‐myristoylated upon caspase‐mediated cleavage and targeted to mitochondria | |
Nakano, Kengo1  Sakurai, Nagisa1  Utsumi, Toshihiko1  Ishisaka, Rumi1  | |
[1] Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan | |
关键词: Posttranslational modification; Posttranslational N-myristoylation; Caspase substrate; Protein N-myristoylation; Protein acylation; Actin; tBID; truncated BID; tActin; truncated actin; TNF; tumor necrosis factor; NMT; N-myristoyltransferase; PCR; polymerase chain reaction; DPBS; Dulbecco's phosphate-buffered saline; SDS; sodium dodecyl sulfate; PAGE; polyacrylamide gel electrophoresis; | |
DOI : 10.1016/S0014-5793(03)00180-7 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
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.
【 授权许可】
Unknown
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