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

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

doi:10.1002/1439-7633(20010803)2:7/8<550::aid-cbic550>3.3.co;2-k

Cited by 33 publications

(65 citation statements)

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“…3 much, as terms containing s c2 are dispersed, while terms containing s c1 are dominated by either s r or s M , because s c1 is so long. The rotational correlation time of calbindin, s r , is around 4·10 -9 s [47]. A contribution of s M to the correlation time is possible.…”

Section: Discussionmentioning

confidence: 99%

A paramagnetic probe to localize residues next to carboxylates on protein surfaces

et al. 2002

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It is shown that the paramagnetic properties of lanthanides can be exploited to obtain information on specific parts of a protein surface. Owing to the high affinity of coordinatively unsaturated lanthanide complexes for oxygen donors, carboxylate groups can be used as preferential targets for the interaction. The DO3A ligand is particularly useful in these studies, as it coordinates lanthanides in a heptadentate fashion, leaving two sites available for exogenous donors. A solution of a (15)N-labeled sample protein, calbindin D(9k) (75 residues), was titrated with up to 200% of Gd(III)-DO3A complex, and an inversion recovery (15)N-(1)H HSQC experiment was used to measure the paramagnetic contributions to the longitudinal relaxation rates of the amide protons. Relaxation data were used as distance constraints to estimate the number of interacting complexes and the occupancies of their binding sites. Four preferential interaction sites on the protein surface are found. Inspection of the various carboxylate side chains on the surface of the protein indicates that Gd(III)-DO3A interacts preferentially with carboxylate-rich regions, rather than with isolated carboxylates, suggesting the possibility of chelation of one Gd(III)-DO3A molecule by two carboxylate groups. Gd(III)-DO3A is thus a valuable semi-selective probe for clusters of negative charges on the protein surface.

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Section: Discussionmentioning

confidence: 99%

A paramagnetic probe to localize residues next to carboxylates on protein surfaces

et al. 2002

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“…Experiments that rely on heteronuclei, in particular on 13 C, were recently applied to several paramagnetic proteins, where the contribution to line broadening from the paramagnetic center is so large that 1 H signals are broadened beyond detectable limits in a wide sphere around the metal ion [10][11][12][13][14]. These applications promoted the revival of heteronuclear NMR as a means to overcome the limitations imposed to 1 H NMR by fast proton transverse relaxation [15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Novel 13C direct detection experiments, including extension to the third dimension, to perform the complete assignment of proteins

Bermel

Bertini

Felli

et al. 2006

Journal of Magnetic Resonance

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“…Thus, by measuring the paramagnetic relaxation enhancements for a sufficient number of protein nuclei, the location of the paramagnetic metal ion can be determined. This has been demonstrated for the calcium-binding protein calbindin D 9k , where the calcium ion was substituted by the paramagnetic lanthanide ion, Ce 3+ [54], and the location of the metal ion in the protein structure was determined from the interactions between the unpaired electrons and the protein nuclei. The R 2p relaxation and, thus, the line broadening observed in paramagnetic metalloproteins can be very severe.…”

Section: Paramagnetic Metal Ionsmentioning

confidence: 99%

Investigating metal-binding in proteins by nuclear magnetic resonance

Jensen

Hass

Hansen

et al. 2007

Cell. Mol. Life Sci.

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Metal ions play a key role for the function of many proteins. The interaction of the metal ion with the protein and its involvement in the function of the protein vary widely. In some proteins, the metal ion is bound tightly to the ligand residues and may be the key player in the function of the protein, as in the case of blue copper proteins. In other proteins, the metal ion is bound only temporarily and loosely to the protein, as in the case of some metalloenzymes and other proteins where the metal ion acts as a cofactor necessary for the function of the protein. Such proteins are often known as metal ion-activated proteins. The review focuses on recent nuclear magnetic resonance (NMR) studies of a series of metal-dependent proteins and the characterization of the metal-binding sites. In particular, we focus on NMR techniques for studying metal binding to proteins such as chemical shift mapping, paramagnetic NMR and changes in backbone dynamics upon metal binding.

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

Cited by 33 publications

References 0 publications

A paramagnetic probe to localize residues next to carboxylates on protein surfaces

A paramagnetic probe to localize residues next to carboxylates on protein surfaces

Novel 13C direct detection experiments, including extension to the third dimension, to perform the complete assignment of proteins

Investigating metal-binding in proteins by nuclear magnetic resonance

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