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Structure-based approaches to create new E2-E3 enzyme pairs.

G Sebastiaan Winkler·H Th Marc Timmers
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Abstract

The study of ubiquitin-conjugating enzymes (E2) and ubiquitin-protein ligases (E3) is complicated by the fact that a relatively limited number of E2 proteins interacts with a large number of E3 enzymes. Many E3 enzymes contain a RING domain. Based on structural and biochemical analysis of the complex between UbcH5b and the CNOT4 RING finger, we describe a rationale to design new E2-E3 enzyme pairs with altered specificity. In such enzyme pairs, the E2 and E3 proteins are each mutated so that they do not interact with their wild-type partner. However, a functional enzyme pair is reconstituted when both E2 and E3 mutants are combined. Such altered-specificity enzyme pairs may be valuable to study the physiological significance of particular E2-E3 interactions.

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Amino Acid SequenceMolecular Sequence DataMutagenesis, Site-DirectedNuclear Magnetic Resonance, BiomolecularProtein ConformationSequence Homology, Amino AcidUbiquitin-Conjugating EnzymesUbiquitin-Protein LigasesJournal ArticleResearch Support, Non-U.S. Gov't

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G Sebastiaan Winkler, H Th Marc Timmers. 2005. Structure-based approaches to create new E2-E3 enzyme pairs.. https://doi.org/10.1016/s0076-6879(05)99024-1

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