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Structural Basis for Non-Covalent Interaction Between Ubiquitin and the Ubiquitin Conjugating Enzyme Variant Human MMS2

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Abstract

Modification of proteins by post-translational covalent attachment of a single, or chain, of ubiquitin molecules serves as a signaling mechanism for a number of regulatory functions in eukaryotic cells. For example, proteins tagged with lysine-63 linked polyubiquitin chains are involved in error-free DNA repair. The catalysis of lysine-63 linked polyubiquitin chains involves the sequential activity of three enzymes (E1, E2, and E3) that ultimately transfer a ubiquitin thiolester intermediate to a protein target. The E2 responsible for catalysis of lysine-63 linked polyubiquitination is a protein heterodimer consisting of a canonical E2 known as Ubc13, and an E2-like protein, or ubiquitin conjugating enzyme variant (UEV), known as Mms2. We have determined the solution structure of the complex formed by human Mms2 and ubiquitin using high resolution, solution state nuclear magnetic resonance (NMR) spectroscopy. The structure of the Mms2–Ub complex provides important insights into the molecular basis underlying the catalysis of lysine-63 linked polyubiquitin chains.

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Abbreviations

Ub:

ubiquitin

E1:

ubiquitin activating enzyme

E2:

ubiquitin conjugating enzyme

E3:

ubiquitin ligase

UEV:

ubiquitin conjugating enzyme variant

CUE:

similar to yeast protein Cue1p

UBA:

ubiquitin associated domain

UIM:

ubiquitin interacting motif

NMR:

nuclear magnetic resonance

ITC:

isothermal titration calorimetry

IPTG:

isopropyl ß-D-thiogalactopyranoside

NOE:

nuclear Overhauser effect

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Authors and Affiliations

  1. Department of Biochemistry, University of Alberta, Edmonton, Alberta, T6G 2H7, Canada

    Michael J. Lewis, Linda F. Saltibus, D. Duong Hau & Leo Spyracopoulos

  2. Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5E5, Canada

    Wei Xiao

Authors
  1. Michael J. Lewis
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  2. Linda F. Saltibus
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  3. D. Duong Hau
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  4. Wei Xiao
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  5. Leo Spyracopoulos
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Correspondence to Leo Spyracopoulos.

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Lewis, M.J., Saltibus, L.F., Hau, D.D. et al. Structural Basis for Non-Covalent Interaction Between Ubiquitin and the Ubiquitin Conjugating Enzyme Variant Human MMS2. J Biomol NMR 34, 89–100 (2006). https://doi.org/10.1007/s10858-005-5583-6

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  • DOI: https://doi.org/10.1007/s10858-005-5583-6

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