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Locating the Metal Ion in Calcium-Binding Proteins by Using Cerium(III) as a Probe

2001; Wiley; Volume: 2; Issue: 7-8 Linguagem: Inglês

10.1002/1439-7633(20010803)2

1439-7633

Ivano Bertini, Yong‐Min Lee, Claudio Luchinat, Mario Piccioli, Luisa Poggi,

Mass Spectrometry Techniques and Applications

ChemBioChemVolume 2, Issue 7-8 p. 550-558 Full Paper Locating the Metal Ion in Calcium-Binding Proteins by Using Cerium(III) as a Probe Ivano Bertini Prof. Dr., Ivano Bertini Prof. Dr. [email protected] Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorYong-Min Lee Dr., Yong-Min Lee Dr. Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorClaudio Luchinat Prof. Dr., Claudio Luchinat Prof. Dr. Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorMario Piccioli Dr., Mario Piccioli Dr. Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorLuisa Poggi, Luisa Poggi Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this author Ivano Bertini Prof. Dr., Ivano Bertini Prof. Dr. [email protected] Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorYong-Min Lee Dr., Yong-Min Lee Dr. Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorClaudio Luchinat Prof. Dr., Claudio Luchinat Prof. Dr. Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorMario Piccioli Dr., Mario Piccioli Dr. Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this authorLuisa Poggi, Luisa Poggi Magnetic Resonance Center (CERM) University of Florence Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy, Fax: (+390) 55-4574271Search for more papers by this author First published: 27 July 2001 https://doi.org/10.1002/1439-7633(20010803)2:7/8 3.0.CO;2-TCitations: 63Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat Graphical Abstract Direct 13C detection (see picture) in combination with paramagnetic versions of classic double- and triple-resonance NMR spectroscopic experiments permits the identification of the ligands bound to the paramagnetic metal ion in the cerium-substituted calcium-binding protein calbindin D9k. This method provides a tool to characterize the coordination sphere of lanthanide ions in metalloproteins in solution. Abstract The detection and assignment of NMR spectroscopic signals of carbon atoms from carbonyl and carboxylate groups in the loop hosting the CeIII ion was performed for the cerium-substituted calcium-binding protein calbindin D9k. This provided a tool to characterize in solution the first coordination sphere of the metal ion. Due to the well-documented possibility of replacing calcium with metal ions of the LnIII series, this approach turns out to be extremely efficient for characterizing in solution the coordination of calcium ions in proteins, independently of the availability of X-ray crystal structures. The present approach completes the structural characterization of lanthanide-substituted calcium-binding proteins, for which the role of long-range constraints arising from hyperfine interaction and self-orientation has already been assessed. References 1 I. Bertini, C. Luchinat, M. Piccioli, Methods Enzymol. 2001, in press. 2 S. Linse, P. Brodin, T. Drakenberg, E. Thulin, P. Sellers, K. Elmden, T. Grundstrom, S. Forsén, Biochemistry 1987, 26, 6723–6735. 3 M. Akke, S. Forsén, W. J. Chazin, J. Mol. Biol. 1991, 220, 173–189. 4 H. J. Vogel, T. Drakenberg, S. Forsén, J. D. O′Neil, T. Hofmann, Biochemistry 1985, 24, 3870–3876. 5 H. M. McConnell, R. E. Robertson, J. Chem. Phys. 1958, 29, 1361–1365. 6 R. J. 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