Apoptosis-associated caspase activation assays

被引：68

作者：

Kaufmann, Scott H. ^{[1
,2
]}

Lee, Sun-Hee ^{[2
]}

Meng, X. Wei ^{[1
,2
]}

Loegering, David A. ^{[1
]}

Kottke, Timothy J. ^{[1
]}

Henzing, Alexander J. ^{[3
]}

Ruchaud, Sandrine ^{[3
]}

Samejima, Kumiko ^{[3
]}

Earnshaw, William C. ^{[3
]}

机构：

[1] Mayo Clin, Div Oncol Res, Coll Med, Rochester, MN 55905 USA

[2] Mayo Clin, Coll Med, Dept Mol Pharmacol & Expt Therapeut, Rochester, MN 55905 USA

[3] Univ Edinburgh, Inst Cell & Mol Biol, Edinburgh EH9 3JR, Midlothian, Scotland

来源：

METHODS | 2008年 / 44卷 / 03期

关键词：

caspases; apoptosis; affinity labeling; enzymatic activity; fluorogenic substrate; flow cytometry; immunoblotting; immunofluorescence;

D O I：

10.1016/j.ymeth.2007.11.005

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Caspases are aspartate-directed cysteine proteases that cleave a diverse group of intracellular substrates to contribute to various manifestations of apoptosis. These proteases are synthesized as inactive precursors and are activated as a consequence of signaling induced by a wide range of physiological and pathological stimuli. Caspase activation can be detected by measurement of catalytic activity, immunoblotting for cleavage of their substrates, immunolabeling using conformation-sensitive antibodies or affinity labeling followed by flow cytometry or ligand blotting. Here we describe methods for each of these assays, identify recent improvements in these assays and outline the strengths and limitations of each approach. (C) 2007 Elsevier Inc. All rights reserved.

引用

页码：262 / 272

页数：11

共 68 条

[11] Multiple species of CPP32 and Mch2 are the major active caspases present in apoptotic cells [J].

Faleiro, L ;

Kobayashi, R ;

Fearnhead, H ;

Lazebnik, Y .

EMBO JOURNAL, 1997, 16 (09) :2271-2281

[12] Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas [J].

Faubion, WA ;

Guicciardi, ME ;

Miyoshi, H ;

Bronk, SF ;

Roberts, PJ ;

Svingen, PA ;

Kaufmann, SH ;

Gores, GJ .

JOURNAL OF CLINICAL INVESTIGATION, 1999, 103 (01) :137-145

[13] In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains [J].

FernandesAlnemri, T ;

Armstrong, RC ;

Krebs, J ;

Srinivasula, SM ;

Wang, L ;

Bullrich, F ;

Fritz, LC ;

Trapani, JA ;

Tomaselli, KJ ;

Litwack, G ;

Alnemri, ES .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (15) :7464-7469

[14]

FERNANDESALNEMRI T, 1995, CANCER RES, V55, P2737

[15] Many cuts to ruin:: a comprehensive update of caspase substrates [J].

Fischer, U ;

Jänicke, RU ;

Schulze-Osthoff, K .

CELL DEATH AND DIFFERENTIATION, 2003, 10 (01) :76-100

[16] Inhibition of human caspases by peptide-based and macromolecular inhibitors [J].

Garcia-Calvo, M ;

Peterson, EP ;

Leiting, B ;

Ruel, R ;

Nicholson, DW ;

Thornberry, NA .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (49) :32608-32613

[17] Cleavage of automodified poly(ADP-ribose) polymerase during apoptosis - Evidence for involvement of caspase-7 [J].

Germain, M ;

Affar, EB ;

D'Amours, D ;

Dixit, VM ;

Salvesen, GS ;

Poirier, GG .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (40) :28379-28384

[18] Caspase-1 activation of caspase-6 in human apoptotic neurons [J].

Guo, H ;

Pétrin, D ;

Zhang, Y ;

Bergeron, C ;

Goodyer, CG ;

LeBlanc, AC .

CELL DEATH AND DIFFERENTIATION, 2006, 13 (02) :285-292

[19] A sequential two-step mechanism for the production of the mature p17:p12 form of caspase-3 in vitro [J].

Han, ZY ;

Hendrickson, EA ;

Bremner, TA ;

Wyche, JH .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (20) :13432-13436

[20]

Hawkins CJ, 2000, J BIOL CHEM, V275, P27084

← 1 2 3 4 5 6 7 →