The latent form of macropain (high molecular weight multicatalytic protease) restores ATP-dependent proteolysis to soluble extracts of BHK fibroblasts pretreated with anti-macropain antibodies

Michael J. McGuire, George N. DeMartino

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Specific immunoadsorption of the high molecular weight multicatalytic protease, macropain, from postmicrosomal extracts of BHK fibroblasts inhibited ATP-dependent proteolysis of exogenous protein substrates. The immunoprecipitated macropain represented the latent (L) form of the protease because it had low protease activity but was activated by methods that activate purified macropain L. Reconstitution of the antibody-treated extracts with purified macropain L, but not macropain A, from bovine heart or human erythrocytes, completely restored ATP-dependent proteolysis, even though ATP did not directly activate either purified macropain L or the immunoprecipitated protease. Reconstituted ATP-dependent proteolysis was saturable with respect to added macropain and never exceeded the level of proteolysis present in the original extract. These results indicate that macropain L plays a key role in ATP-dependent proteolysis but suggest that the protease may require interaction with or modification by another cellular component to demonstrate this effect.

Original languageEnglish (US)
Pages (from-to)911-916
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume160
Issue number2
DOIs
StatePublished - Apr 28 1989

Fingerprint

ATP-Dependent Proteases
Proteolysis
Proteasome Endopeptidase Complex
Fibroblasts
Anti-Idiotypic Antibodies
Peptide Hydrolases
Adenosine Triphosphate
Molecular Weight
Molecular weight
Antibodies
L Forms
Erythrocytes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

@article{25a02d990aa54f7d94cdced58e3bf1af,
title = "The latent form of macropain (high molecular weight multicatalytic protease) restores ATP-dependent proteolysis to soluble extracts of BHK fibroblasts pretreated with anti-macropain antibodies",
abstract = "Specific immunoadsorption of the high molecular weight multicatalytic protease, macropain, from postmicrosomal extracts of BHK fibroblasts inhibited ATP-dependent proteolysis of exogenous protein substrates. The immunoprecipitated macropain represented the latent (L) form of the protease because it had low protease activity but was activated by methods that activate purified macropain L. Reconstitution of the antibody-treated extracts with purified macropain L, but not macropain A, from bovine heart or human erythrocytes, completely restored ATP-dependent proteolysis, even though ATP did not directly activate either purified macropain L or the immunoprecipitated protease. Reconstituted ATP-dependent proteolysis was saturable with respect to added macropain and never exceeded the level of proteolysis present in the original extract. These results indicate that macropain L plays a key role in ATP-dependent proteolysis but suggest that the protease may require interaction with or modification by another cellular component to demonstrate this effect.",
author = "McGuire, {Michael J.} and DeMartino, {George N.}",
year = "1989",
month = "4",
day = "28",
doi = "10.1016/0006-291X(89)92521-7",
language = "English (US)",
volume = "160",
pages = "911--916",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - The latent form of macropain (high molecular weight multicatalytic protease) restores ATP-dependent proteolysis to soluble extracts of BHK fibroblasts pretreated with anti-macropain antibodies

AU - McGuire, Michael J.

AU - DeMartino, George N.

PY - 1989/4/28

Y1 - 1989/4/28

N2 - Specific immunoadsorption of the high molecular weight multicatalytic protease, macropain, from postmicrosomal extracts of BHK fibroblasts inhibited ATP-dependent proteolysis of exogenous protein substrates. The immunoprecipitated macropain represented the latent (L) form of the protease because it had low protease activity but was activated by methods that activate purified macropain L. Reconstitution of the antibody-treated extracts with purified macropain L, but not macropain A, from bovine heart or human erythrocytes, completely restored ATP-dependent proteolysis, even though ATP did not directly activate either purified macropain L or the immunoprecipitated protease. Reconstituted ATP-dependent proteolysis was saturable with respect to added macropain and never exceeded the level of proteolysis present in the original extract. These results indicate that macropain L plays a key role in ATP-dependent proteolysis but suggest that the protease may require interaction with or modification by another cellular component to demonstrate this effect.

AB - Specific immunoadsorption of the high molecular weight multicatalytic protease, macropain, from postmicrosomal extracts of BHK fibroblasts inhibited ATP-dependent proteolysis of exogenous protein substrates. The immunoprecipitated macropain represented the latent (L) form of the protease because it had low protease activity but was activated by methods that activate purified macropain L. Reconstitution of the antibody-treated extracts with purified macropain L, but not macropain A, from bovine heart or human erythrocytes, completely restored ATP-dependent proteolysis, even though ATP did not directly activate either purified macropain L or the immunoprecipitated protease. Reconstituted ATP-dependent proteolysis was saturable with respect to added macropain and never exceeded the level of proteolysis present in the original extract. These results indicate that macropain L plays a key role in ATP-dependent proteolysis but suggest that the protease may require interaction with or modification by another cellular component to demonstrate this effect.

UR - http://www.scopus.com/inward/record.url?scp=0024980075&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024980075&partnerID=8YFLogxK

U2 - 10.1016/0006-291X(89)92521-7

DO - 10.1016/0006-291X(89)92521-7

M3 - Article

C2 - 2719706

AN - SCOPUS:0024980075

VL - 160

SP - 911

EP - 916

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -