Purification and characterization of a multiprotein component of the Drosophila 26 S (1500 kDa) proteolytic complex

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Abstract

A multiprotein complex, referred to as the μ particle, was purified to apparent homogeneity from Drosophila melanogaster embryos. This multiprotein complex has no protease activity, but it can be incorporated into an even larger multiprotein complex which exhibits strong and selective protease activity, i.e. it degrades only ubiquitin-conjugated proteins. Incorporation of the μ particle into the ubiquitin conjugate-degrading larger complex is absolutely ATP-dependent. On these criteria the larger complex corresponds to the 26 S (1500 kDa) proteolytic complex partially purified and characterized from reticulocytes. A procedure is described for the purification of the Drosophila 26 S (1500 kDa) proteolytic complex. It was found to be a stoichiometric complex of the μ particle and the 20 S proteosome. Although no other polypeptide was present in stoichiometric amount in the 26 S (1500 kDa) proteolytic complex besides the μ particle and the 20 S proteosome, an additional protein factor(s) is required for its assembly, the ubiquitin conjugate-degrading activity cannot be reconstituted from the purified μ particle and the 20 S proteosome. Synthesis of the μ particle is developmentally regulated; its concentration is highest in embryos. This is probably connected with massive degradation of yolk proteins during embryogenesis. In chicken, rabbit and human cells a high molecular weight multiprotein complex can be detected, which is immunologically related to the Drosophila μ particle.

Original languageEnglish
Pages (from-to)9055-9062
Number of pages8
JournalJournal of Biological Chemistry
Volume268
Issue number12
Publication statusPublished - Apr 25 1993

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Multiprotein Complexes
Drosophila
Purification
Ubiquitin
Peptide Hydrolases
Embryonic Structures
Egg Proteins
Reticulocytes
Drosophila melanogaster
Embryonic Development
Chickens
Proteins
Adenosine Triphosphate
Molecular Weight
Molecular weight
Cells
Rabbits
Degradation
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

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abstract = "A multiprotein complex, referred to as the μ particle, was purified to apparent homogeneity from Drosophila melanogaster embryos. This multiprotein complex has no protease activity, but it can be incorporated into an even larger multiprotein complex which exhibits strong and selective protease activity, i.e. it degrades only ubiquitin-conjugated proteins. Incorporation of the μ particle into the ubiquitin conjugate-degrading larger complex is absolutely ATP-dependent. On these criteria the larger complex corresponds to the 26 S (1500 kDa) proteolytic complex partially purified and characterized from reticulocytes. A procedure is described for the purification of the Drosophila 26 S (1500 kDa) proteolytic complex. It was found to be a stoichiometric complex of the μ particle and the 20 S proteosome. Although no other polypeptide was present in stoichiometric amount in the 26 S (1500 kDa) proteolytic complex besides the μ particle and the 20 S proteosome, an additional protein factor(s) is required for its assembly, the ubiquitin conjugate-degrading activity cannot be reconstituted from the purified μ particle and the 20 S proteosome. Synthesis of the μ particle is developmentally regulated; its concentration is highest in embryos. This is probably connected with massive degradation of yolk proteins during embryogenesis. In chicken, rabbit and human cells a high molecular weight multiprotein complex can be detected, which is immunologically related to the Drosophila μ particle.",
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AB - A multiprotein complex, referred to as the μ particle, was purified to apparent homogeneity from Drosophila melanogaster embryos. This multiprotein complex has no protease activity, but it can be incorporated into an even larger multiprotein complex which exhibits strong and selective protease activity, i.e. it degrades only ubiquitin-conjugated proteins. Incorporation of the μ particle into the ubiquitin conjugate-degrading larger complex is absolutely ATP-dependent. On these criteria the larger complex corresponds to the 26 S (1500 kDa) proteolytic complex partially purified and characterized from reticulocytes. A procedure is described for the purification of the Drosophila 26 S (1500 kDa) proteolytic complex. It was found to be a stoichiometric complex of the μ particle and the 20 S proteosome. Although no other polypeptide was present in stoichiometric amount in the 26 S (1500 kDa) proteolytic complex besides the μ particle and the 20 S proteosome, an additional protein factor(s) is required for its assembly, the ubiquitin conjugate-degrading activity cannot be reconstituted from the purified μ particle and the 20 S proteosome. Synthesis of the μ particle is developmentally regulated; its concentration is highest in embryos. This is probably connected with massive degradation of yolk proteins during embryogenesis. In chicken, rabbit and human cells a high molecular weight multiprotein complex can be detected, which is immunologically related to the Drosophila μ particle.

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