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Guerra-Castellano A, Díaz-Quintana A, Moreno-Beltrán B, et al. Mimicking Tyrosine Phosphorylation in Human Cytochrome c by the Evolved tRNA Synthetase Technique. Chemistry. 2015;21(42):15004-12doi: 10.1002/chem.201502019.
Guerra-Castellano, A., Díaz-Quintana, A., Moreno-Beltrán, B., López-Prados, J., Nieto, P. M., Meister, W., Staffa, J., Teixeira, M., Hildebrandt, P., De la Rosa, M. A., & Díaz-Moreno, I. (2015). Mimicking Tyrosine Phosphorylation in Human Cytochrome c by the Evolved tRNA Synthetase Technique. Chemistry (Weinheim an der Bergstrasse, Germany), 21(42), 15004-12. https://doi.org/10.1002/chem.201502019
Guerra-Castellano, Alejandra, et al. "Mimicking Tyrosine Phosphorylation in Human Cytochrome c by the Evolved tRNA Synthetase Technique." Chemistry (Weinheim an der Bergstrasse, Germany) vol. 21,42 (2015): 15004-12. doi: https://doi.org/10.1002/chem.201502019
Guerra-Castellano A, Díaz-Quintana A, Moreno-Beltrán B, López-Prados J, Nieto PM, Meister W, Staffa J, Teixeira M, Hildebrandt P, De la Rosa MA, Díaz-Moreno I. Mimicking Tyrosine Phosphorylation in Human Cytochrome c by the Evolved tRNA Synthetase Technique. Chemistry. 2015 Oct 12;21(42):15004-12. doi: 10.1002/chem.201502019. Epub 2015 Aug 21. PMID: 26329855.
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Chemistry. 2015 Oct 12;21(42):15004-12. doi: 10.1002/chem.201502019. Epub 2015 Aug 21.

Mimicking Tyrosine Phosphorylation in Human Cytochrome c by the Evolved tRNA Synthetase Technique.

Chemistry (Weinheim an der Bergstrasse, Germany)

Alejandra Guerra-Castellano, Antonio Díaz-Quintana, Blas Moreno-Beltrán, Javier López-Prados, Pedro M Nieto, Wiebke Meister, Jana Staffa, Miguel Teixeira, Peter Hildebrandt, Miguel A De la Rosa, Irene Díaz-Moreno

Affiliations

  1. IBVF - cicCartuja, Universidad de Sevilla - CSIC, Avenida Américo Vespucio 49, Sevilla 41092 (Spain).
  2. IBVF - cicCartuja, Universidad de Sevilla - CSIC, Avenida Américo Vespucio 49, Sevilla 41092 (Spain). [email protected].
  3. IIQ - cicCartuja, Universidad de Sevilla - CSIC, Avenida Américo Vespucio 49, Sevilla 41092 (Spain).
  4. Technische Universität Berlin, Institut für Chemie, Sekr. PC14, Strasse des 17. Juni 135, 10623 Berlin (Germany).
  5. Instituto de Tecnologia Química e Biológica, António Xavier Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras (Portugal).
  6. IBVF - cicCartuja, Universidad de Sevilla - CSIC, Avenida Américo Vespucio 49, Sevilla 41092 (Spain). [email protected].

PMID: 26329855 DOI: 10.1002/chem.201502019

Abstract

Phosphorylation of tyrosine 48 of cytochrome c is related to a wide range of human diseases due to the pleiotropic role of the heme-protein in cell life and death. However, the structural conformation and physicochemical properties of phosphorylated cytochrome c are difficult to study as its yield from cell extracts is very low and its kinase remains unknown. Herein, we report a high-yielding synthesis of a close mimic of phosphorylated cytochrome c, developed by optimization of the synthesis of the non-canonical amino acid p-carboxymethyl-L-phenylalanine (pCMF) and its efficient site-specific incorporation at position 48. It is noteworthy that the Y48pCMF mutation significantly destabilizes the Fe-Met bond in the ferric form of cytochrome c, thereby lowering the pKa value for the alkaline transition of the heme-protein. This finding reveals the differential ability of the phosphomimic protein to drive certain events. This modified cytochrome c might be an important tool to investigate the role of the natural protein following phosphorylation.

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keywords: NMR spectroscopy; cytochromes; heme proteins; phosphorylation; protein modifications

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