TY - JOUR
T1 - Cleavage of lamin a by Mch2α but not CPP32
T2 - Multiple interleukin 1β-con verting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis
AU - Takahashi, Atsushi
AU - Alnemri, Emad S.
AU - Lazebnik, Yuri A.
AU - Fernandes-Alnemri, Teresa
AU - Litwack, Gerald
AU - Moir, Robert D.
AU - Goldman, Robert D.
AU - Poirier, Guy G.
AU - Kaufmann, Scott H.
AU - Earnshaw, William C.
PY - 1996/8/6
Y1 - 1996/8/6
N2 - Although proteases related to the interleukin 1β-converting enzyme (ICE) are known to be essential for apoptotic execution, the number of enzymes involved, their substrate specificities, and their specific roles in the characteristic biochemical and morphological changes of apoptosis are currently unknown. These questions were addressed using cloned recombinant ICE-related proteases (IRPs) and a cell-free model system for apoptosis (S/M extracts). First, we compared the substrate specificities of two recombinant human IRPs, CPP32 and Mch2α. Both enzymes cleaved poly-(ADP-ribose) polymerase, albeit with different efficiencies. Mch2α also cleaved recombinant and nuclear lamin A at a conserved VEIDNG sequence located in the middle of the coiled-coil rod domain, producing a fragment that was indistinguishable from the lamin A fragment observed in S/M extracts and in apoptotic cells. In contrast, CPP32 did not cleave lamin A. The cleavage of lamin A by Mch2a and by S/M extracts was inhibited by millimolar concentrations of Zn2+, which had a minimal effect on cleavage of poly(ADP-ribose) polymerase by CPP32 and by S/M extracts. We also found that n-(acetyltyrosinyrvalinyL-Nε-biotinyllysyl)aspartic acid [(2, 6-dimethylbenzoyl)oxy] methyl ketone, which derivatizes the larger subunit of active ICE, can affinity label up to five active IRPs in S/M extracts. Together, these observations indicate that the processing of nuclear proteins in apoptosis involves multiple IRPs having distinct preferences for their apoptosis-associated substrates.
AB - Although proteases related to the interleukin 1β-converting enzyme (ICE) are known to be essential for apoptotic execution, the number of enzymes involved, their substrate specificities, and their specific roles in the characteristic biochemical and morphological changes of apoptosis are currently unknown. These questions were addressed using cloned recombinant ICE-related proteases (IRPs) and a cell-free model system for apoptosis (S/M extracts). First, we compared the substrate specificities of two recombinant human IRPs, CPP32 and Mch2α. Both enzymes cleaved poly-(ADP-ribose) polymerase, albeit with different efficiencies. Mch2α also cleaved recombinant and nuclear lamin A at a conserved VEIDNG sequence located in the middle of the coiled-coil rod domain, producing a fragment that was indistinguishable from the lamin A fragment observed in S/M extracts and in apoptotic cells. In contrast, CPP32 did not cleave lamin A. The cleavage of lamin A by Mch2a and by S/M extracts was inhibited by millimolar concentrations of Zn2+, which had a minimal effect on cleavage of poly(ADP-ribose) polymerase by CPP32 and by S/M extracts. We also found that n-(acetyltyrosinyrvalinyL-Nε-biotinyllysyl)aspartic acid [(2, 6-dimethylbenzoyl)oxy] methyl ketone, which derivatizes the larger subunit of active ICE, can affinity label up to five active IRPs in S/M extracts. Together, these observations indicate that the processing of nuclear proteins in apoptosis involves multiple IRPs having distinct preferences for their apoptosis-associated substrates.
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U2 - 10.1073/pnas.93.16.8395
DO - 10.1073/pnas.93.16.8395
M3 - Article
C2 - 8710882
AN - SCOPUS:0029758833
SN - 0027-8424
VL - 93
SP - 8395
EP - 8400
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 16
ER -