Multiple forms of ubiquitin-protein ligase. Binding of activated ubiquitin to protein substrates

Pauline L. Lee, Christian F. Midelfort, Koko Murakami, Victor Bernard Hatcher

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Enzyme activities that catalyzed the covalent attachment of ubiquitin to protein substrates (ubiquitin-protein ligase, UbL) were purified from the extracts of human red blood cells. These activities required the presence of ubiquitin-activating enzyme and ATP for activity. Four fractions (UbL A, B1, B2, and C) were resolved and showed different specificities toward added substrates [carboxymethylated bovine serum albumin (CM-BSA), G-actin, lysozyme, and α-lactalbumin]. The enzyme fractions gave different products with a given substrate. UbL A and UbL B1 were exclusively active with CM-BSA and α-lactalbumin, respectively. UbL B2 was most active toward CM-BSA with substantial activities to G-actin and α-lactalbumin and with no activity to lysozyme. UbL C showed significant activities with all four substrates, having a highest activity toward CM-BSA. There were many endogenous proteins present in the erythrocyte extract which were efficient substrates for ubiquitin conjugation. In particular, a pair of substrates were identified from eythrocyte extracts which were far more efficient substrates than the denatured proteins exogenously added.

Original languageEnglish (US)
Pages (from-to)3134-3138
Number of pages5
JournalBiochemistry
Volume25
Issue number11
StatePublished - 1986

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Ubiquitin-Protein Ligases
Ubiquitin
Bovine Serum Albumin
Lactalbumin
Substrates
Proteins
Muramidase
Actins
Ubiquitin-Activating Enzymes
Erythrocytes
Enzymes
Protein C
Enzyme activity
Adenosine Triphosphate
Blood
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Multiple forms of ubiquitin-protein ligase. Binding of activated ubiquitin to protein substrates. / Lee, Pauline L.; Midelfort, Christian F.; Murakami, Koko; Hatcher, Victor Bernard.

In: Biochemistry, Vol. 25, No. 11, 1986, p. 3134-3138.

Research output: Contribution to journalArticle

Lee, Pauline L. ; Midelfort, Christian F. ; Murakami, Koko ; Hatcher, Victor Bernard. / Multiple forms of ubiquitin-protein ligase. Binding of activated ubiquitin to protein substrates. In: Biochemistry. 1986 ; Vol. 25, No. 11. pp. 3134-3138.
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