TAILIEUCHUNG - Báo cáo khoa học: A simple protocol to study blue copper proteins by NMR

In the case of oxidized plastocyanin fromSynechocystissp. PCC6803, an NMR approach based on classical two and three dimensional experiments for sequential assignment leaves unobserved 14 out of 98 amino acids. A protocol which simply makes use of tailored versions of 2D HSQC and 3D CBCA(CO)NH and CBCANH leads to the identi-fication of nine of the above 14 residues. The proposed protocol differs from previous aproaches in that it does not involve the use of unconventional experiments designed specifically for paramagnetic systems, and does not exploit the occurrence of a corresponding diamagnetic species in chemical exchange with the blue copper form. . | Eur. J. Biochem. 270 600-609 2003 FEBS 2003 doi A simple protocol to study blue copper proteins by NMR loannis Gelis1. Nikolaos Katsaros1. Claudio Luchinat2 3 Mario Piccioli2 4 and Luisa Poaai2 4 1NCSR Demokritos Institute of Physical Chemistry Agia Paraskevi Attikis Greece 2Magnetic Resonance Center 3 Department of Agricultural Biotechnology and 4 Department of Chemistry University of Florence Italy In the case of oxidized plastocyanin from Synechocystis sp. PCC6803 an NMR approach based on classical two and three dimensional experiments for sequential assignment leaves unobserved 14 out of 98 amino acids. A protocol which simply makes use of tailored versions of 2D HSQC and 3D CBCA CO NH and CBCANH leads to the identification of nine of the above 14 residues. The proposed protocol differs from previous aproaches in that it does not involve the use of unconventional experiments designed specifically for paramagnetic systems and does not exploit the occurrence of a corresponding diamagnetic species in chemical exchange with the blue copper form. This protocol is expected to extend the popularity of NMR in the structural studies of copper II proteins allowing researchers to increase the amount of information available via NMR on the neighborhood of a paramagnetic center without requiring a specific expertise in the field. The resulting 3D spectra are standard spectra that can be handled by any standard software for protein NMR data analysis. Keywords blue copper proteins NMR spectroscopy structural biology paramagnetic proteins plastocyanin. There is a strong interest in the structural biology community for the study of copper trafficking and copper homeostasis 1-11 . This involves the understanding of the role of metal ions in protein folding and misfolding related diseases 12-20 as well as the understanding at the atomic level of protein-protein interactions in electron-transfer processes 21-29 . Within this framework the search .

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