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. 2017 May;7(3):390-394.
doi: 10.1016/j.apsb.2016.12.008. Epub 2017 Feb 28.

Structural analysis of recombinant human ubiquitin-conjugating enzyme UbcH5c

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Structural analysis of recombinant human ubiquitin-conjugating enzyme UbcH5c

Fangshu Wu et al. Acta Pharm Sin B. 2017 May.

Abstract

UbcH5c belongs to the ubiquitin-conjugating enzyme family and plays an important role in catalyzing ubiquitination during TNF-α--triggered NF-κB activation. Therefore, UbcH5c is a potent therapeutic target for the treatment of inflammatory and autoimmune diseases induced by aberrant activation of NF-κB. In this study, we established a stable expression system for recombinant UbcH5c and solved the crystal structure of UbcH5c belonging to space group P22121 with one molecule in the asymmetric unit. This study provides the basis for further study of UbcH5c including the design of UbcH5c inhibitors.

Keywords: Crystal structure; Inflammatory target; NF-κB; UbcH5c; Ubiquitin-conjugating enzyme; Ubiquitination.

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Figures

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Graphical abstract
Fig. 1
Figure 1
(A) Size exclusion chromatography for the analysis of UbcH5c; (B) SDS-PAGE analysis of purification of UbcH5c.
Fig. 2
Figure 2
One crystal of UbcH5c grown at 293 K (100×).
Fig. 3
Figure 3
Overall view of the structure of UbcH5c. (A) Cartoon representation of the UbcH5c. The N and C termini of UbcH5c are labeled N and C. Secondary structure elements of UbcH5c are labeled α (cyan), β (magenta) and L (pink). The active site (C85) is shown as sticks. (B) Secondary structure of the UbcH5c is shown as the amino acid sequence. Bars (cyan) indicate helix and arrows (magenta) indicate β-strand.
Fig. 4
Figure 4
Superimposition of the structures between the PDB entry 5egg (magenta) and the chain A of PDB entry 1x23 (cyan). The active site (C85) is shown as sticks.
Fig. 5
Figure 5
Details of the active site environment in the crystal structure of UbcH5c.

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