Inhibition of the calcium(2+) ion-dependent ATPase from sarcoplasmic reticulum by dicyclohexylcarbodiimide: evidence for location of the calcium(2+) ion binding site in a hydrophobic region

Biochemistry ◽  
1979 ◽  
Vol 18 (1) ◽  
pp. 108-113 ◽  
Author(s):  
Uri Pick ◽  
Efraim Racker
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Binding Site ◽  
Ion Binding ◽  
Hydrophobic Region ◽  
Dependent Atpase

scholarly journals Functional characterization of the dimerization domain of the ferric uptake regulator (Fur) of Pseudomonas aeruginosa

2006 ◽  
Vol 400 (3) ◽  
pp. 385-392 ◽  
Author(s):  
Erdeni Bai ◽  
Federico I. Rosell ◽  
Bao Lige ◽  
Marcia R. Mauk ◽  
Barbara Lelj-Garolla ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Metal Ions ◽  
Binding Site ◽  
Metal Ion ◽  
Association Constants ◽  
Ion Binding ◽  
Ferric Uptake Regulator ◽  
Metal Ion Binding ◽  
Dimerization Domain ◽  
High Affinity

The functional properties of the recombinant C-terminal dimerization domain of the Pseudomonas aeruginosa Fur (ferric uptake regulator) protein expressed in and purified from Escherichia coli have been evaluated. Sedimentation velocity measurements demonstrate that this domain is dimeric, and the UV CD spectrum is consistent with a secondary structure similar to that observed for the corresponding region of the crystallographically characterized wild-type protein. The thermal stability of the domain as determined by CD spectroscopy decreases significantly as pH is increased and increases significantly as metal ions are added. Potentiometric titrations (pH 6.5) establish that the domain possesses a high-affinity and a low-affinity binding site for metal ions. The high-affinity (sensory) binding site demonstrates association constants (KA) of 10(±7)×106, 5.7(±3)×106, 2.0(±2)×106 and 2.0(±3)×104 M−1 for Ni2+, Zn2+, Co2+ and Mn2+ respectively, while the low-affinity (structural) site exhibits association constants of 1.3(±2)×106, 3.2(±2)×104, 1.76(±1)×105 and 1.5(±2)×103 M−1 respectively for the same metal ions (pH 6.5, 300 mM NaCl, 25 °C). The stability of metal ion binding to the sensory site follows the Irving–Williams order, while metal ion binding to the partial sensory site present in the domain does not. Fluorescence experiments indicate that the quenching resulting from binding of Co2+ is reversed by subsequent titration with Zn2+. We conclude that the domain is a reasonable model for many properties of the full-length protein and is amenable to some analyses that the limited solubility of the full-length protein prevents.

Location of N-cyclohexyl-N'-(4-dimethyl-amino-alpha-naphthyl)carbodiimide-binding site in sarcoplasmic reticulum Ca2+-transporting ATPase

1998 ◽  
Vol 253 (1) ◽  
pp. 339-344 ◽  
Author(s):  
Isabel Velasco-Guillen ◽  
Senena Corbalan-Garcia ◽  
Juan C. Gomez-Fernandez ◽  
Jose A. Teruel
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Binding Site

An Antibody to Annexin V Blocks Ca2+-Dependent ATPase Activity of Sarcoplasmic Reticulum in Human Failing Hearts

1998 ◽  
Vol 853 (1 CARDIAC SARCO) ◽  
pp. 333-337 ◽  
Author(s):  
ION I. MORARU ◽  
SERGEI SYRBU ◽  
NADA ZECEVIC ◽  
DAVID W. HAGER ◽  
JAMES WATRAS ◽  
...  
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Atpase Activity ◽  
Annexin V ◽  
Dependent Atpase
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