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Volumn 281, Issue 5381, 1998, Pages 1355-1357

Essential role of CED-4 oligomerization in CED-3 activation and apoptosis

Author keywords

[No Author keywords available]

Indexed keywords

CASPASE; CED 4 PROTEIN; CED 9 PROTEIN; CYSTEINE PROTEINASE; PROTEIN BCL 2; UNCLASSIFIED DRUG;

EID: 0032575723     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.281.5381.1355     Document Type: Article
Times cited : (242)

References (27)
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    • note
    • Each FLAG-CED-4 mutant was cotransfected with AU1-CED-3(C358S) into 293T cells. Cell lysates were immunoprecipitated with antibody to AU1 (anti-AU1) (Babco) and immunoblotted with anti-FLAG. Each CED-4 mutant interacted with CED-3(C358S) as well as wild-type CED-4.
  • 24
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    • note
    • L, and Apaf-1(1-465) were each fused with a COOH-terminal FLAG epitope tag in pRK5. CED-3(C358S) was also fused to a COOH-terminal AU1 epitope tag in pRK5. Myc - CED-4 and Myc - CED-4 (K165R) were described (3). Myc - Apaf-1(1-465) and Myc - CED-3(C358S) were made in pcDNA3.1(-)/ MycHis (Invitrogen) with a COOH-terminal Myc tag. Authenticity of each construct was confirmed by DNA sequencing.
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    • 3543003591 scopus 로고    scopus 로고
    • note
    • 6 293T cells [grown in Dulbecco's minimal essential medium supplemented with 10% fetal bovine serum, penicillin-streptomycin (100 U/ml), and glutamine (1 mM)] were plated per 60-mm dish. Twenty-four hours after transfection by the calcium phosphate method, cells were lysed in 300 μl of IP-lysis buffer [50 mM Hepes (pH 7.4), 1% NP-40, 150 mM NaCl, 10% glycerol, 1 mM EDTA, 2 mM dithiothreitol] supplemented with 1 mM phenylmethylsulfonyl fluoride and 1% aprotinin. Extracts (100 μl) were diluted 1:1 in IP-lysis buffer and immunoprecipitated with antibody for 3 hours at 4°C, washed with 600 μl of IP-lysis buffer, and resolved by SDS - polyacrylamide gel electrophoresis (SDS-PAGE).
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    • X. Yang et al., Cell 89, 1067 (1997).
    • (1997) Cell , vol.89 , pp. 1067
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    • 3543023396 scopus 로고    scopus 로고
    • note
    • We thank F. Chen and H. R. Horvitz for reagents and discussions, P. Svec and A. Darbinian for technical support, and V. M. Dixit, M. Gilman, S. L. Schreiber, and X. Wang for reagents. Supported by the Leukemia Society of America (to X.Y.), the Medical Scientist Training Program (to H.Y.C.), and NIH grant CA51462.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.