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Semi-Synthesis of Labeled Proteins for Spectroscopic Applications
Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, Napoli 80134, Italy
Dipartimento delle Scienze Biologiche, Università di Napoli "Federico II", Via Mezzocannone 16, Napoli 80134, Italy
* Author to whom correspondence should be addressed.
Received: 28 November 2012; in revised form: 21 December 2012 / Accepted: 24 December 2012 / Published: 2 January 2013
Abstract: Since the introduction of SPPS by Merrifield in the 60s, peptide chemists have considered the possibility of preparing large proteins. The introduction of native chemical ligation in the 90s and then of expressed protein ligation have opened the way to the preparation of synthetic proteins without size limitations. This review focuses on semi-synthetic strategies useful to prepare proteins decorated with spectroscopic probes, like fluorescent labels and stable isotopes, and their biophysical applications. We show that expressed protein ligation, combining the advantages of organic chemistry with the easy and size limitless recombinant protein expression, is an excellent strategy for the chemical synthesis of labeled proteins, enabling a single protein to be functionalized at one or even more distinct positions with different probes.
Keywords: expressed protein ligation; intein; Förster resonance energy transfer; segmental labeling; NMR; protein labeling
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De Rosa, L.; Russomanno, A.; Romanelli, A.; D'Andrea, L.D. Semi-Synthesis of Labeled Proteins for Spectroscopic Applications. Molecules 2013, 18, 440-465.
De Rosa L, Russomanno A, Romanelli A, D'Andrea LD. Semi-Synthesis of Labeled Proteins for Spectroscopic Applications. Molecules. 2013; 18(1):440-465.
De Rosa, Lucia; Russomanno, Anna; Romanelli, Alessandra; D'Andrea, Luca D. 2013. "Semi-Synthesis of Labeled Proteins for Spectroscopic Applications." Molecules 18, no. 1: 440-465.