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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Direct characterization of E2-dependent target specificity and processivity using an artificial p27-linker-E2 ubiquitination system

  • Ryu, Kyoung-Seok (Magnetic Resonance Team, Korea Basic Science Institute) ;
  • Choi, Yun-Seok (Magnetic Resonance Team, Korea Basic Science Institute) ;
  • Ko, Jun-Sang (Department of Chemistry and National Creative Research Initiative Center, KAIST) ;
  • Kim, Seong-Ock (Department of Chemistry and National Creative Research Initiative Center, KAIST) ;
  • Kim, Hyun-Jung (Magnetic Resonance Team, Korea Basic Science Institute) ;
  • Cheong, Hae-Kap (Magnetic Resonance Team, Korea Basic Science Institute) ;
  • Jeon, Young-Ho (Magnetic Resonance Team, Korea Basic Science Institute) ;
  • Choi, Byong-Seok (Department of Chemistry and National Creative Research Initiative Center, KAIST) ;
  • Cheong, Chae-Joon (Magnetic Resonance Team, Korea Basic Science Institute)
  • Published : 2008.12.31
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Abstract

Little attention has been paid to the specificity between E2 and the target protein during ubiquitination, although RING-E3 induces a potential intra-molecular reaction by mediating the direct transfer of ubiquitin from E2 to the target protein. We have constructed artificial E2 fusion proteins in which a target protein (p27) is tethered to one of six E2s via a flexible linker. Interestingly, only three E2s (UbcH5b, hHR6b, and Cdc34) are able to ubiquitinate p27 via an intra-molecular reaction in this system. Although the first ubiquitination of p27 (p27-Ub) by Cdc34 is less efficient than that of UbcH5b and hHR6b, the additional ubiquitin attachment to p27-Ub by Cdc34 is highly efficient. The E2 core of Cdc34 provides specificity to p27, and the residues 184-196 are required for possessive ubiquitination by Cdc34. We demonstrate direct E2 specificity for p27 and also show that differential ubiquitin linkages can be dependent on E2 alone.

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References

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