Conformational changes in the 20S proteasome upon macromolecular ligand binding analyzed with monoclonal antibodies

Arch Biochem Biophys. 1999 Feb 15;362(2):325-8. doi: 10.1006/abbi.1998.1037.

Abstract

Proteasomes interact with a variety of macromolecular ligands that modulate their ability to degrade peptide and protein substrates. The effector PA28 increases the peptidase activities of proteasomes whereas HSP90 and alpha-crystallin inhibit a peptide-hydrolyzing activity. Four monoclonal antibodies were used as probes to detect conformational changes of proteasome subunits. Conformational changes in alpha- or beta-subunits were found upon binding PA28, HSP90, alpha-crystallin, and the substrate casein but not with the peptide substrate analogs calpain inhibitor 1 (Ac-Leu-Leu-norleucinal), calpain inhibitor 2 (Ac-Leu-Leu-methioninal), or MG 132 (N-Cbz-Leu-Leu-leucinal).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Affinity
  • Binding Sites, Antibody
  • Caseins / metabolism
  • Crystallins / metabolism
  • Cysteine Endopeptidases / chemistry*
  • Cysteine Endopeptidases / immunology
  • Cysteine Endopeptidases / metabolism*
  • Cysteine Proteinase Inhibitors / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Glycoproteins / metabolism
  • HSP90 Heat-Shock Proteins / metabolism
  • Humans
  • Leupeptins / metabolism
  • Ligands
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / immunology
  • Multienzyme Complexes / metabolism*
  • Muscle Proteins*
  • Oligopeptides / metabolism
  • Proteasome Endopeptidase Complex
  • Protein Conformation
  • Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Caseins
  • Crystallins
  • Cysteine Proteinase Inhibitors
  • Glycoproteins
  • HSP90 Heat-Shock Proteins
  • Leupeptins
  • Ligands
  • Multienzyme Complexes
  • Muscle Proteins
  • Oligopeptides
  • PSME1 protein, human
  • Proteins
  • calpain inhibitors
  • calpain inhibitor 2
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde