z-logo
Premium
Characterization of functionally independent domains in the human ubiquitin conjugating enzyme UbcH2
Author(s) -
Kaiser Peter,
Mandl Sonja,
Schweiger Manfred,
Schneider Rainer
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01323-7
Subject(s) - ubiquitin , ubiquitin conjugating enzyme , ubiquitins , enzyme , biochemistry , histone , deubiquitinating enzyme , biology , mutant , ubiquitin ligase , in vitro , microbiology and biotechnology , chemistry , dna , gene
UbcH2 encodes a human ubiquitin conjugating enzyme (E2) able to conjugate ubiquitin to histone H2A in an E3 independent manner in vitro, which indicates that UbcH2 directly interacts with its substrates. To identify parts of the enzyme that are capable of binding H2A, we expressed several deletion mutants of UbcH2 in E. coli and tested the ability of the affinity purified mutant proteins to ubiquitinate H2A in the presence of bacterial expressed E1 and ubiquitin. With this in vitro assay we identified a C‐terminal part of UbcH2 to be important for the interaction with H2A. Transfer of this C‐terminal domain to another human E2, which is unable to catalyze ubiquitination of histones, leads to a fully active hybrid human ubiquitin conjugating enzyme capable of H2A ubiquitination. These results demonstrate that UbcH2 consists of two functionally independent domains. A N‐terminal core domain with ubiquitin conjugating activity, and a C‐terminal domain which interacts with substrate proteins.

This content is not available in your region!

Continue researching here.

Having issues? You can contact us here