Dual-Functional-Tag-Facilitated Protein Labeling and Immobilization

ACS Omega
Xinyi ZhangYinan Wei

Abstract

An important strategy in the construction of biomimetic membranes and devices is to use natural proteins as the functional components for incorporation in a polymeric or nanocomposite matrix. Toward this goal, an important step is to immobilize proteins with high efficiency and precision without disrupting the protein function. Here, we developed a dual-functional tag containing histidine and the non-natural amino acid azidohomoalanine (AHA). AHA is metabolically incorporated into the protein, taking advantage of the Met-tRNA and Met-tRNA synthetase. Histidine in the tag can facilitate metal-affinity purification, whereas AHA can react with an alkyne-functionalized probe or surface via well-established click chemistry. We tested the performance of the tag using two model proteins, green fluorescence protein and an enzyme pyrophosphatase. We found that the addition of the tag and the incorporation of AHA did not significantly impair the properties of these proteins, and the histidine-AHA tag can facilitate protein purification, immobilization, and labeling.

References

Dec 1, 1985·Molecular and Cellular Biology·G I EvanJ M Bishop
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·A Einhauer, A Jungbauer
Dec 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·Kristi L KiickCarolyn R Bertozzi
Aug 1, 2002·Journal of the American Chemical Society·Jason W ChinPeter G Schultz
Oct 14, 2005·Journal of the American Chemical Society·A James Link, David A Tirrell
Dec 22, 2005·Nature Biotechnology·Jean-Denis PédelacqGeoffrey S Waldo
Dec 19, 2006·ACS Chemical Biology·Nicholas J AgardCarolyn R Bertozzi
Apr 24, 2007·Nature Chemical Biology·Isaac S CarricoCarolyn R Bertozzi
Jul 21, 2009·Journal of the American Chemical Society·John M AntosHidde L Ploegh
Mar 20, 2010·Journal of the American Chemical Society·Bijan Zakeri, Mark Howarth
Jun 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Chayasith UttamapinantAlice Y Ting
Jun 15, 2010·Nature Neuroscience·Daniela C DieterichErin M Schuman
May 24, 2011·Methods in Molecular Biology·Gertjan KramerChris G de Koster
Mar 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Bijan ZakeriMark Howarth
Apr 25, 2013·Chemical Communications : Chem Comm·Shohei UchinomiyaItaru Hamachi
Aug 24, 2013·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Pramod K AvtiAshok Kakkar
Nov 5, 2013·Molecular Cancer Therapeutics·Manuel SimonUwe Zangemeister-Wittke
Dec 18, 2013·Bioconjugate Chemistry·Jianpeng WangJiang Xia
Jan 18, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ying MaMartino Luigi di Salvo
Jan 30, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Julianne Martell, Eranthie Weerapana
Mar 19, 2014·Chemical Science·Neel H Shah, Tom W Muir
Jan 13, 2015·Journal of Structural Biology·Chathurada S GajadeeraOleg V Tsodikov
Feb 26, 2015·Proceedings of the National Academy of Sciences of the United States of America·Yau-Tsz LaiHongzhe Sun
Mar 17, 2015·Nature Methods·Susanne tom DieckErin M Schuman
Oct 8, 2015·Journal of Proteome Research·Daniel B McClatchyJohn R Yates
Oct 31, 2015·Current Opinion in Chemical Biology·Samuel C Reddington, Mark Howarth
Nov 20, 2016·Chembiochem : a European Journal of Chemical Biology·Matthias P ExnerNediljko Budisa

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Citations

Jun 28, 2018·Bioconjugate Chemistry·Maksymilian Marek ZegotaTanja Weil

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Methods Mentioned

BETA
Fluorescence
protein folding
Electrophoresis

Software Mentioned

SigmaPlot

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