Abstract:
:We have previously reported that calpastatin, an endogenous inhibitory protein of calpain, is cleaved by a caspase-3-like protease during apoptosis in human Jurkat T cells [Kato, M. et al. (2000) J. Biochem. 127, 297-305]. In this study, we found that nonmuscle myosin heavy chain-A (NMHC-A) is cleaved during apoptosis in Jurkat cells by using a cleavage-site-directed antibody for calpastatin. The cleavage-site-directed antibody was raised against the amino-terminal fragment of calpastatin, and this antibody detected the in vitro cleaved calpastatin fragment. Although cleaved calpastatin was not detected, a 95-kDa polypeptide (p95) was detected in apoptotic cells by this antibody. This p95 was identified as the carboxyl-terminal fragment of NMHC-A based on the results of peptide mass spectrometry fingerprinting and amino-terminal sequencing. Furthermore, two cleavage sites on NMHC-A, Asp-1153 and Asp-1948, were determined, and three cleaved fragments of NMHC-A, one cleaved at Asp-1153 and the other two cleaved at Asp-1948, were detected by cleavage-site-directed antibodies against each cleavage site. The results of confocal immunofluorescence microscopic analysis show that the cleavage at Asp-1948 occurs faster than that at Asp-1153 during apoptosis. In addition, the Asp-1153 cleaved fragment was distributed diffusely in the cytoplasm of apoptotic cells, whereas the Asp-1948 cleaved fragments were detected as condensed dots. In conclusion, our findings can be summarized as follows: (i) NMHC-A is cleaved at two sites during apoptosis, (ii) the timing of cleavage is different between these two cleavage sites, and (iii) the distribution of cleaved fragments is different in apoptotic cells.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Kato M,Fukuda H,Nonaka T,Imajoh-Ohmi Sdoi
10.1093/jb/mvi015keywords:
subject
Has Abstractpub_date
2005-02-01 00:00:00pages
157-66issue
2eissn
0021-924Xissn
1756-2651pii
137/2/157journal_volume
137pub_type
杂志文章abstract::Using the methods of methylation/GLC, MS, and 1H-NMR analyses, a disaccharide structure of GalNAc alpha 1----2Fuc1---- was identified as one of the major structural units at non-reducing terminals of a highly branched and N-glycosidically linked carbohydrate on an allergenically active glycopeptide, Gp-2 (Mr ca. 8,000...
journal_title:Journal of biochemistry
pub_type: 杂志文章
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doi:10.1093/oxfordjournals.jbchem.a131657
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a133463
更新日期:1981-07-01 00:00:00
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pub_type: 杂志文章
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doi:10.1093/oxfordjournals.jbchem.a124018
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1978-10-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1997-09-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a133972
更新日期:1982-09-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134298
更新日期:1983-06-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a122419
更新日期:1988-07-01 00:00:00
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pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a130824
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123092
更新日期:1990-04-01 00:00:00
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pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124455
更新日期:1994-06-01 00:00:00
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pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021254
更新日期:1996-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a130916
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pub_type: 杂志文章
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pub_type: 杂志文章
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:2009-07-01 00:00:00
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pub_type: 杂志文章
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更新日期:2007-07-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a135333
更新日期:1985-09-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1985-08-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a121919
更新日期:1987-02-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1980-10-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1987-10-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123863
更新日期:1992-07-01 00:00:00