Characterization of Tescalcin, a Novel EF-Hand Protein with a Single Ca2+-Binding Site:  Metal-Binding Properties, Localization in Tissues and Cells, and Effect on Calcineurin†

Biochemistry ◽  
2003 ◽  
Vol 42 (49) ◽  
pp. 14553-14565 ◽  
Author(s):  
Christina Gutierrez-Ford ◽  
Konstantin Levay ◽  
Aldrin V. Gomes ◽  
Erasmo M. Perera ◽  
Tapan Som ◽  
...  
Keyword(s):  
Binding Site ◽  
Metal Binding ◽  
Binding Properties ◽  
Ef Hand

scholarly journals Binding of Gd3+to the neuronal signalling protein calexcitin identifies an exchangeable Ca2+-binding site

2016 ◽  
Vol 72 (4) ◽  
pp. 276-281
Author(s):  
Lucas Chataigner ◽  
Jingxu Guo ◽  
Peter T. Erskine ◽  
Alun R. Coker ◽  
Steve P. Wood ◽  
...  
Keyword(s):  
Metal Ions ◽  
Binding Site ◽  
Metal Binding ◽  
Calcium Signalling ◽  
Specific Protein ◽  
Metal Binding Sites ◽  
Calcium Sensing ◽  
Neuronal Signalling ◽  
Ef Hand ◽  
Ef Hands

Calexcitin was first identified in the marine snailHermissenda crassicornisas a neuronal-specific protein that becomes upregulated and phosphorylated in associative learning. Calexcitin possesses four EF-hand motifs, but only the first three (EF-1 to EF-3) are involved in binding metal ions. Past work has indicated that under physiological conditions EF-1 and EF-2 bind Mg2+and Ca2+, while EF-3 is likely to bind only Ca2+. The fourth EF-hand is nonfunctional owing to a lack of key metal-binding residues. The aim of this study was to use a crystallographic approach to determine which of the three metal-binding sites of calexcitin is most readily replaced by exogenous metal ions, potentially shedding light on which of the EF-hands play a `sensory' role in neuronal calcium signalling. By co-crystallizing recombinant calexcitin with equimolar Gd3+in the presence of trace Ca2+, EF-1 was shown to become fully occupied by Gd3+ions, while the other two sites remain fully occupied by Ca2+. The structure of the Gd3+–calexcitin complex has been refined to anRfactor of 21.5% and anRfreeof 30.4% at 2.2 Å resolution. These findings suggest that EF-1 of calexcitin is the Ca2+-binding site with the lowest selectivity for Ca2+, and the implications of this finding for calcium sensing in neuronal signalling pathways are discussed.

scholarly journals Purification of a Variant-specific Surface Protein ofGiardia lambliaand Characterization of its Metal-binding Properties

1995 ◽  
Vol 270 (23) ◽  
pp. 13807-13813 ◽  
Author(s):  
Hugo D. Luján ◽  
Michael R. Mowatt ◽  
Jing-jing Wu ◽  
Yun Lu ◽  
Andrew Lees ◽  
...  
Keyword(s):  
Specific Surface ◽  
Metal Binding ◽  
Surface Protein ◽  
Binding Properties

scholarly journals Characterization of the DNA- and Metal-Binding Properties of Vibrio anguillarum Fur Reveals Conservation of a Structural Zn2+ Ion

2000 ◽  
Vol 182 (21) ◽  
pp. 6264-6267 ◽  
Author(s):  
Ekaterina E. Zheleznova ◽  
Jorge H. Crosa ◽  
Richard G. Brennan
Keyword(s):  
Metal Binding ◽  
Iron Uptake ◽  
Vibrio Anguillarum ◽  
Zinc Ion ◽  
Ferric Uptake Regulator ◽  
Binding Properties ◽  
E Coli ◽  
Solution Studies

ABSTRACT The ferric uptake regulator, Fur, represses iron uptake and siderophore biosynthetic genes under iron-replete conditions. Here we report in vitro solution studies on Vibrio anguillarum Fur binding to the consensus 19-bp Escherichia coli iron box in the presence of several divalent metals. We found that V. anguillarum Fur binds the iron box in the presence of Mn2+, Co2+, Cd2+, and to a lesser extent Ni2+ but, unlike E. coli Fur, not in the presence of Zn2+. We also found that V. anguillarum Fur contains a structural zinc ion that is necessary yet alone is insufficient for DNA binding.

Characterization of Ligands of a High-Affinity Metal-Binding Site in the Latent Chloroplast F1-ATPase by EPR Spectroscopy of Bound VO2+

Biochemistry ◽  
1994 ◽  
Vol 33 (16) ◽  
pp. 4910-4917 ◽  
Author(s):  
Andrew L. P. Houseman ◽  
Lola Morgan ◽  
Russell LoBrutto ◽  
Wayne D. Frasch
Keyword(s):  
Binding Site ◽  
Epr Spectroscopy ◽  
Metal Binding ◽  
Metal Binding Site ◽  
F1 Atpase ◽  
High Affinity

scholarly journals Unravelling the Structure of the Tetrahedral Metal-Binding Site in METP3 through an Experimental and Computational Approach

Molecules ◽  
2021 ◽  
Vol 26 (17) ◽  
pp. 5221
Author(s):  
Salvatore La Gatta ◽  
Linda Leone ◽  
Ornella Maglio ◽  
Maria De Fenza ◽  
Flavia Nastri ◽  
...  
Keyword(s):  
Metal Ions ◽  
Binding Site ◽  
Metal Binding ◽  
Binding Sites ◽  
Metal Ion ◽  
Structural Determinants ◽  
Binding Properties ◽  
Tetrahedral Geometry ◽  
Design Synthesis ◽  
Metal Binding Sites

Understanding the structural determinants for metal ion coordination in metalloproteins is a fundamental issue for designing metal binding sites with predetermined geometry and activity. In order to achieve this, we report in this paper the design, synthesis and metal binding properties of METP3, a homodimer made up of a small peptide, which self assembles in the presence of tetrahedrally coordinating metal ions. METP3 was obtained through a redesign approach, starting from the previously developed METP molecule. The undecapeptide sequence of METP, which dimerizes to house a Cys4 tetrahedral binding site, was redesigned in order to accommodate a Cys2His2 site. The binding properties of METP3 were determined toward different metal ions. Successful assembly of METP3 with Co(II), Zn(II) and Cd(II), in the expected 2:1 stoichiometry and tetrahedral geometry was proven by UV-visible spectroscopy. CD measurements on both the free and metal-bound forms revealed that the metal coordination drives the peptide chain to fold into a turned conformation. Finally, NMR data of the Zn(II)-METP3 complex, together with a retrostructural analysis of the Cys-X-X-His motif in metalloproteins, allowed us to define the model structure. All the results establish the suitability of the short METP sequence for accommodating tetrahedral metal binding sites, regardless of the first coordination ligands.

scholarly journals Effect of Common Buffers and Heterocyclic Ligands on the Binding of Cu(II) at the Multimetal Binding Site in Human Serum Albumin

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Magdalena Sokołowska ◽  
Krystyna Pawlas ◽  
Wojciech Bal
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Binding Site ◽  
Metal Binding ◽  
Visible Range ◽  
Heterocyclic Ligands ◽  
Binding Properties ◽  
Spectral Changes ◽  
Conformational Effects

Visible-range circular dichroism titrations were used to study Cu(II) binding properties of Multimetal Binding Site (MBS) of Human Serum Albumin (HSA). The formation of ternary MBS-Cu(II)-Buffer complexes at pH 7.4 was positively verified for sodium phosphate, Tris, and Hepes, the three most common biochemical buffers. The phosphate > Hepes > Tris order of affinities, together with strong spectral changes induced specifically by Tris, indicates the presence of both Buffer-Cu(II) and Buffer-HSA interactions. All complexes are strong enough to yield a nearly 100% ternary complex formation in 0.5 mM HSA dissolved in 100 mM solutions of respective buffers. The effects of warfarin and ibuprofen, specific ligands of hydrophobic pockets I and II in HSA on the Cu(II) binding to MBS were also investigated. The effects of ibuprofen were negligible, but warfarin diminished the MBS affinity for Cu(II) by a factor of 20, as a result of indirect conformational effects. These results indicate that metal binding properties of MBS can be modulated directly and indirectly by small molecules.

scholarly journals Characterization of the Metal-Binding Site in Aminopeptidase B

2006 ◽  
Vol 29 (12) ◽  
pp. 2378-2382 ◽  
Author(s):  
Junzo Hirose ◽  
Takamichi Ohsaki ◽  
Naoyo Nishimoto ◽  
Shouji Matuoka ◽  
Takashi Hiromoto ◽  
...  
Keyword(s):  
Binding Site ◽  
Metal Binding ◽  
Metal Binding Site
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