Cleavage of lamin a by Mch2α but not CPP32: Multiple interleukin 1β-con verting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis

Atsushi Takahashi, Emad S. Alnemri, Yuri A. Lazebnik, Teresa Fernandes-Alnemri, Gerald Litwack, Robert D. Moir, Robert D. Goldman, Guy G. Poirier, Scott H Kaufmann, William C. Earnshaw

Research output: Contribution to journalArticle

460 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)8395-8400
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number16
StatePublished - Aug 6 1996

Fingerprint

Lamins
Caspase 1
Interleukin-1
Lamin Type A
Peptide Hydrolases
Apoptosis
Enzymes
Poly(ADP-ribose) Polymerases
Substrate Specificity
Affinity Labels
Cell-Free System
Conserved Sequence
Nuclear Proteins
Ketones
Aspartic Acid

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Cleavage of lamin a by Mch2α but not CPP32 : Multiple interleukin 1β-con verting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis. / Takahashi, Atsushi; Alnemri, Emad S.; Lazebnik, Yuri A.; Fernandes-Alnemri, Teresa; Litwack, Gerald; Moir, Robert D.; Goldman, Robert D.; Poirier, Guy G.; Kaufmann, Scott H; Earnshaw, William C.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, No. 16, 06.08.1996, p. 8395-8400.

Research output: Contribution to journalArticle

Takahashi, Atsushi ; Alnemri, Emad S. ; Lazebnik, Yuri A. ; Fernandes-Alnemri, Teresa ; Litwack, Gerald ; Moir, Robert D. ; Goldman, Robert D. ; Poirier, Guy G. ; Kaufmann, Scott H ; Earnshaw, William C. / Cleavage of lamin a by Mch2α but not CPP32 : Multiple interleukin 1β-con verting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis. In: Proceedings of the National Academy of Sciences of the United States of America. 1996 ; Vol. 93, No. 16. pp. 8395-8400.
@article{81327a0ad0a544f0aa203fe8d19d6b9e,
title = "Cleavage of lamin a by Mch2α but not CPP32: Multiple interleukin 1β-con verting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis",
abstract = "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.",
author = "Atsushi Takahashi and Alnemri, {Emad S.} and Lazebnik, {Yuri A.} and Teresa Fernandes-Alnemri and Gerald Litwack and Moir, {Robert D.} and Goldman, {Robert D.} and Poirier, {Guy G.} and Kaufmann, {Scott H} and Earnshaw, {William C.}",
year = "1996",
month = "8",
day = "6",
language = "English (US)",
volume = "93",
pages = "8395--8400",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "16",

}

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.

UR - http://www.scopus.com/inward/record.url?scp=0029758833&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029758833&partnerID=8YFLogxK

M3 - Article

C2 - 8710882

AN - SCOPUS:0029758833

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

SN - 0027-8424

IS - 16

ER -