| http://purl.uniprot.org/citations/18359862 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/18359862 | http://www.w3.org/2000/01/rdf-schema#comment | "We have studied the binding of Zn2+ to the hexa EF-hand protein, calbindin D(28k)-a strong Ca2+-binder involved in apoptosis regulation-which is highly expressed in brain tissue. By use of radioblots, isothermal titration calorimetry, and competition with a fluorescent Zn2+ chelator, we find that calbindin D(28k) binds Zn2+ to three rather strong sites with dissociation constants in the low micromolar range. Furthermore, we conclude based on spectroscopic investigations that the Zn2+-bound state is structurally distinct from the Ca2+-bound state and that the two forms are incompatible, yielding negative allosteric interaction between the zinc- and calcium-binding events. ANS titrations reveal a change in hydrophobicity upon binding Zn2+. The binding of Zn2+ is compatible with the ability of calbindin to activate myo-inositol monophosphatase, one of the known targets of calbindin. Through site-directed mutagenesis, we address the role of cysteine and histidine residues in the binding of Zn2+. Mutation of all five cysteines into serines has no effect on Zn2+-binding affinity or stoichiometry. However, mutating histidine 80 into a glutamine reduces the binding affinity of the strongest Zn2+ site, indicating that this residue is involved in coordinating the Zn2+ ion in this site. Mutating histidines 5, 22, or 114 has significantly smaller effects on Zn2+-binding affinity."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.org/dc/terms/identifier | "doi:10.1110/ps.073381108"xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/author | "Nilsson H."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/author | "Linse S."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/author | "Malm J."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/author | "Thulin E."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/author | "Frohm B."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/author | "Bauer M.C."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/date | "2008"xsd:gYear |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/name | "Protein Sci"xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/pages | "760-767"xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/title | "Zn2+ binding to human calbindin D(28k) and the role of histidine residues."xsd:string |
| http://purl.uniprot.org/citations/18359862 | http://purl.uniprot.org/core/volume | "17"xsd:string |
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