Copper activation of superoxide dismutase 1 (SOD1) in vivo

Role for protein-protein interactions with the copper chaperone for SOD1

Paul J. Schmidt, Catherine Kunst, Valeria L Culotta

Research output: Contribution to journalArticle

Abstract

Insertion of copper into superoxide dismutase 1 (SOD1) in vivo requires the copper chaperone for SOD1 (CCS). CCS encompasses three protein domains: copper binding Domains I and III at the amino and carboxyl termini, and a central Domain II homologous to SOD1. Using a yeast interaction mating system, yeast CCS was seen to physically interact with SOD1, and this interaction required sequences at the predicted dimer interface of CCS Domain II. Interactions with SOD1 also required sequences of Domain III, but not Domain I. Mutations were introduced at the dimer interface of yeast SOD1, and the corresponding mutant failed to interact with CCS. When loaded with copper independent of CCS, this mutant SOD1 exhibited superoxide scavenging activity, but was normally inactive in vivo because CCS failed to recognize the enzyme. Activation of SOD1 by CCS was also examined using an in vivo assay for copper incorporation into SOD1. Yeast CCS was observed to insert copper into a pre-existing pool of apoSOD1 without the need for new SOD1 synthesis or for protein unfolding by the major SSA cytosolic heat shock proteins. Our data are consistent with a model in which prefolded dimers of apoSOD1 serve as substrate for the CCS copper chaperone.

Original languageEnglish (US)
Pages (from-to)33771-33776
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number43
DOIs
StatePublished - Oct 27 2000

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Superoxide Dismutase
Copper
Chemical activation
Proteins
Yeast
Yeasts
Dimers
Superoxide Dismutase-1
Protein Unfolding
Scavenging
Heat-Shock Proteins
Superoxides
Assays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Copper activation of superoxide dismutase 1 (SOD1) in vivo : Role for protein-protein interactions with the copper chaperone for SOD1. / Schmidt, Paul J.; Kunst, Catherine; Culotta, Valeria L.

In: Journal of Biological Chemistry, Vol. 275, No. 43, 27.10.2000, p. 33771-33776.

Research output: Contribution to journalArticle

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abstract = "Insertion of copper into superoxide dismutase 1 (SOD1) in vivo requires the copper chaperone for SOD1 (CCS). CCS encompasses three protein domains: copper binding Domains I and III at the amino and carboxyl termini, and a central Domain II homologous to SOD1. Using a yeast interaction mating system, yeast CCS was seen to physically interact with SOD1, and this interaction required sequences at the predicted dimer interface of CCS Domain II. Interactions with SOD1 also required sequences of Domain III, but not Domain I. Mutations were introduced at the dimer interface of yeast SOD1, and the corresponding mutant failed to interact with CCS. When loaded with copper independent of CCS, this mutant SOD1 exhibited superoxide scavenging activity, but was normally inactive in vivo because CCS failed to recognize the enzyme. Activation of SOD1 by CCS was also examined using an in vivo assay for copper incorporation into SOD1. Yeast CCS was observed to insert copper into a pre-existing pool of apoSOD1 without the need for new SOD1 synthesis or for protein unfolding by the major SSA cytosolic heat shock proteins. Our data are consistent with a model in which prefolded dimers of apoSOD1 serve as substrate for the CCS copper chaperone.",
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