Structure-based approaches to create new E2-E3 enzyme pairs.
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.
Winkler
G Sebastiaan
GS
Department of Physiological Chemistry, University Medical Center--Utrecht, Utrecht, The Netherlands.
Timmers
H Th Marc
HT
D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry*
D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases