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Cells 2014, 3(2), 639-656; doi:10.3390/cells3020639
Review

The Challenge of Producing Ubiquitinated Proteins for Structural Studies

1
 and 2,*
Received: 4 May 2014; in revised form: 27 May 2014 / Accepted: 28 May 2014 / Published: 12 June 2014
(This article belongs to the Special Issue Protein Ubiquitination)
View Full-Text   |   Download PDF [638 KB, uploaded 12 June 2014]
Abstract: Protein ubiquitination is an important post-translational modification involved in several essential signalling pathways. It has different effects on the target protein substrate, i.e., it can trigger the degradation of the protein in the proteasome, change the interactions of the modified protein with its partners, or affect its localization and activity. In order to understand the molecular mechanisms underlying the consequences of protein ubiquitination, scientists have to face the challenging task of producing ubiquitinated proteins for structural characterization with X-ray crystallography and/or nuclear magnetic resonance (NMR) spectroscopy. These techniques require milligrams of homogeneous samples of high purity. The strategies proposed so far for the production of ubiquitinated proteins can be divided into two groups, i.e., chemical (or non-enzymatic) and enzymatic methodologies. In this review, we summarize the still very sparse examples available in the literature that describe successful production of ubiquitinated proteins amenable for biochemical and structural studies, and discuss advantages and disadvantages of the techniques proposed. We also give a perspective of the direction in which the field might evolve.
Keywords: ubiquitin; post-translational modification; mono-ubiquitination; isopeptide bond; native chemical ligation; non-enzymatic ubiquitination; enzymatic ubiquitination; in vitro ubiquitination; X-ray crystallography; nuclear magnetic resonance spectroscopy ubiquitin; post-translational modification; mono-ubiquitination; isopeptide bond; native chemical ligation; non-enzymatic ubiquitination; enzymatic ubiquitination; in vitro ubiquitination; X-ray crystallography; nuclear magnetic resonance spectroscopy
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Faggiano, S.; Pastore, A. The Challenge of Producing Ubiquitinated Proteins for Structural Studies. Cells 2014, 3, 639-656.

AMA Style

Faggiano S, Pastore A. The Challenge of Producing Ubiquitinated Proteins for Structural Studies. Cells. 2014; 3(2):639-656.

Chicago/Turabian Style

Faggiano, Serena; Pastore, Annalisa. 2014. "The Challenge of Producing Ubiquitinated Proteins for Structural Studies." Cells 3, no. 2: 639-656.



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