A conserved catalytic residue in the ubiquitin-conjugating enzyme family

Pei Ying Wu, Mary Hanlon, Michael Eddins, Colleen Tsui, Richard S. Rogers, Jane P. Jensen, Michael J Matunis, Allan M. Weissman, Cynthia Wolberger, Cecile M. Pickart

Research output: Contribution to journalArticle

Abstract

Ubiquitin (Ub) regulates diverse functions in eukaryotes through its attachment to other proteins. The defining step in this protein modification pathway is the attack of a substrate lysine residue on Ub bound through its C-terminus to the active site cysteine residue of a Ub-conjugating enzyme (E2) or certain Ub ligases (E3s). So far, these E2 and E3 cysteine residues are the only enzyme groups known to participate in the catalysis of conjugation. Here we show that a strictly conserved E2 asparagine residue is critical for catalysis of E2- and E2/RING E3-dependent isopeptide bond formation, but dispensable for upstream and downstream reactions of Ub thiol ester formation. In constrast, the strictly conserved histidine and proline residues immediately upstream of the asparagine are dispensable for catalysis of isopeptide bond formation. We propose that the conserved asparagine side chain stabilizes the oxyanion intermediate formed during lysine attack. The E2 asparagine is the first non-covalent catalytic group to be proposed in any Ub conjugation factor.

Original languageEnglish (US)
Pages (from-to)5241-5250
Number of pages10
JournalThe EMBO journal
Volume22
Issue number19
DOIs
StatePublished - Oct 1 2003

Fingerprint

Ubiquitin-Conjugating Enzymes
Ubiquitin
Asparagine
Catalysis
Lysine
Cysteine
Ligases
Eukaryota
Histidine
Sulfhydryl Compounds
Proline
Catalytic Domain
Esters
Proteins
Substrates
Enzymes

Keywords

  • Catalytic mechanism
  • E2
  • E3
  • Isopeptide
  • Ubiquitin

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Wu, P. Y., Hanlon, M., Eddins, M., Tsui, C., Rogers, R. S., Jensen, J. P., ... Pickart, C. M. (2003). A conserved catalytic residue in the ubiquitin-conjugating enzyme family. The EMBO journal, 22(19), 5241-5250. https://doi.org/10.1093/emboj/cdg501

A conserved catalytic residue in the ubiquitin-conjugating enzyme family. / Wu, Pei Ying; Hanlon, Mary; Eddins, Michael; Tsui, Colleen; Rogers, Richard S.; Jensen, Jane P.; Matunis, Michael J; Weissman, Allan M.; Wolberger, Cynthia; Pickart, Cecile M.

In: The EMBO journal, Vol. 22, No. 19, 01.10.2003, p. 5241-5250.

Research output: Contribution to journalArticle

Wu, PY, Hanlon, M, Eddins, M, Tsui, C, Rogers, RS, Jensen, JP, Matunis, MJ, Weissman, AM, Wolberger, C & Pickart, CM 2003, 'A conserved catalytic residue in the ubiquitin-conjugating enzyme family', The EMBO journal, vol. 22, no. 19, pp. 5241-5250. https://doi.org/10.1093/emboj/cdg501
Wu PY, Hanlon M, Eddins M, Tsui C, Rogers RS, Jensen JP et al. A conserved catalytic residue in the ubiquitin-conjugating enzyme family. The EMBO journal. 2003 Oct 1;22(19):5241-5250. https://doi.org/10.1093/emboj/cdg501
Wu, Pei Ying ; Hanlon, Mary ; Eddins, Michael ; Tsui, Colleen ; Rogers, Richard S. ; Jensen, Jane P. ; Matunis, Michael J ; Weissman, Allan M. ; Wolberger, Cynthia ; Pickart, Cecile M. / A conserved catalytic residue in the ubiquitin-conjugating enzyme family. In: The EMBO journal. 2003 ; Vol. 22, No. 19. pp. 5241-5250.
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