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

The Challenge of Producing Ubiquitinated Proteins for Structural Studies

1email and 2,* email
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)
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.
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
PDF Full-text Download PDF Full-Text [638 KB, uploaded 12 June 2014 14:42 CEST]

Export to BibTeX |

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.

Cells EISSN 2073-4409 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert