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Protein ligation: an enabling technology for the biophysical analysis of proteins

Nature Methods volume 3, pages 429–438 (2006)Cite this article

Abstract

Biophysical techniques such as fluorescence spectroscopy and nuclear magnetic resonance (NMR) spectroscopy provide a window into the inner workings of proteins. These approaches make use of probes that can either be naturally present within the protein or introduced through a labeling procedure. In general, the more control one has over the type, location and number of probes in a protein, then the more information one can extract from a given biophysical analysis. Recently, two related approaches have emerged that allow proteins to be labeled with a broad range of physical probes. Expressed protein ligation (EPL) and protein trans-splicing (PTS) are both intein-based approaches that permit the assembly of a protein from smaller synthetic and/or recombinant pieces. Here we provide some guidelines for the use of EPL and PTS, and highlight how the dovetailing of these new protein chemistry methods with standard biophysical techniques has improved our ability to interrogate protein function, structure and folding.

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Figure 1: Principle of expressed protein ligation (EPL).
Figure 2: Principle of protein trans-splicing (PTS).
Figure 3: Selected applications of protein ligation technologies.

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Acknowledgements

Some of the work discussed in this review was performed by the authors and was supported by the US National Institutes of Health.

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  1. Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, 10021, New York, USA

    Vasant Muralidharan & Tom W Muir

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Muralidharan, V., Muir, T. Protein ligation: an enabling technology for the biophysical analysis of proteins. Nat Methods 3, 429–438 (2006). https://doi.org/10.1038/nmeth886

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