Structure of the divalent metal ion activator binding site of S-adenosylmethionine synthetase studied by vanadyl(IV) electron paramagnetic resonance

Biochemistry ◽  
1984 ◽  
Vol 23 (3) ◽  
pp. 470-478 ◽  
Author(s):  
George D. Markham
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Binding Site ◽  
Metal Ion ◽  
Divalent Metal ◽  
Paramagnetic Resonance ◽  
Divalent Metal Ion ◽  
Adenosylmethionine Synthetase ◽  
Electron Paramagnetic

scholarly journals Electron-paramagnetic-resonance studies on nitrogenase of Klebsiella pneumoniae. Evidence for acetylene- and ethylene-nitrogenase transient complexes

1978 ◽  
Vol 173 (1) ◽  
pp. 277-290 ◽  
Author(s):  
D J Lowe ◽  
R R Eady ◽  
R N F Thorneley
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Klebsiella Pneumoniae ◽  
Binding Site ◽  
Dissociation Constants ◽  
Paramagnetic Resonance ◽  
Fe Protein ◽  
Low Concentrations ◽  
Direct Binding ◽  
Electron Paramagnetic ◽  
Single Binding Site

Klebsiella pneumoniae nitrogenase exhibited four new electron-paramagnetic-resonance signals during turnover at 10 degrees C, pH7.4, which were assigned to intermediates present in low concentrations in the steady state. 57Fe-substituted Mo–Fe protein showed that they arose from Fe–S clusters in the Mo–Fe protein of nitrogenase. The new signals are designated: Ic, g values at 4.67, 3.37 and approx. 2.0; VI, g values at 2.125, 2.000 and 2.000; VII, g values at 5.7 and 5.4; VIII, g values at 2.092, 1.974 and 1.933. The sharp axial signal VI arises from a Fe4S4 cluster at the −1 oxidation level. This signal was only detected in the presence of ethylene and provides the first evidence of an enzyme–product complex for nitrogenase. [13C]Acetylene and [13C]ethylene provided no evidence for direct binding of this substrate and product to the Fe–S clusters giving rise to these signals. The dependence of signal intensities on acetylene concentration indicated two types of binding site, with apparent dissociation constants K less than 16 micron and K approximately 13mM. A single binding site for ethylene (K=1.5mM) was detected. A scheme is proposed for the mechanism of reduction of acetylene to ethylene and inhibition of this reaction by CO.

Dense-shell glycodendrimers: UV/Vis and electron paramagnetic resonance study of metal ion complexation

2009 ◽  
Vol 466 (2117) ◽  
pp. 1489-1513 ◽  
Author(s):  
Dietmar Appelhans ◽  
Ulrich Oertel ◽  
Roberto Mazzeo ◽  
Hartmut Komber ◽  
Jan Hoffmann ◽  
...  
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Metal Ion ◽  
Electron Paramagnetic Resonance Study ◽  
Copper Metabolism ◽  
Paramagnetic Resonance ◽  
Fifth Generation ◽  
Vis Spectroscopy ◽  
Dense Shell ◽  
Biological Environment ◽  
Electron Paramagnetic

The development of dendritic metal ion carrier systems for use in a biological environment is a challenging task as the carrier system must possess multiple features (e.g. a protective shell for metal decomplexation, targeting functions, metal–intradendrimer complexes, etc.) to substitute for the function of metal proteins in processes such as copper metabolism. Thus, Cu(II) complexation by a series of poly(propyleneimine) glycodendrimers ranging up to the fifth generation that have either a dense maltose or maltotriose shell was investigated by UV/Vis spectroscopy and electron paramagnetic resonance (EPR). As a necessary step towards potential biological application, we elucidated the complexation capacity, location of the Cu(II)–dendrimer complexes and the Cu(II) coordination sphere in the dendritic environment. A generation-dependent Cu(II) complexation was found. Furthermore, analysis of the EPR spectra revealed that internal and external Cu(II) coordination and the symmetry (axial and rhombic) of the generated complexes depend on the oligosaccharide shell, dendrimer generation and the relative concentrations of Cu(II) and the dendrimers. Both axial and rhombic symmetries are generation dependent, but also distort with increasing generation number. External coordination of Cu(II) is supported by sugar groups and water molecules. Finally, a third-generation dendrimer with a maltose shell was used to explore the general complexation behaviour of the dendritic poly(propyleneimine) scaffold towards different metal ions [Cu(II), Ag(I), VO(IV), Ni(II), Eu(III) and UO 2 (VI)].

Activation of Bovine Factor X in the Presence of Calcium, Magnesium, Barium or Manganese ion

1978 ◽  
Vol 40 (02) ◽  
pp. 358-367 ◽  
Author(s):  
Robert H Yue ◽  
Menard M Gertler
Keyword(s):  
Metal Ions ◽  
Binding Site ◽  
Metal Ion ◽  
Factor Xa ◽  
Divalent Metal ◽  
Activation Process ◽  
Factor X ◽  
Divalent Metal Ion ◽  
Russell’S Viper ◽  
Bovine Factor

SummaryThe binding of divalent metal ions to bovine factor X, factor Xa and the coagulant protein in Russell’s viper venom was studied by the technique of fluorescence quenching. Titration of factor X with Ca+2, Mg+2 or Ba+2 revealed that these metal ions can bind to factor X. A tightly binding site(s) was observed with Kd of 79 and 98 μM for Ca+2 and Mg+2 respectively. A loosely binding site(s) was evident with Kd of 0.55, 0.50 and 0.35 mM for Ca+2, Mg+2 and Ba+2 respectively. The quenching phenomenon was also observed when Mn+2 was used as titrant but factor X precipitated out when the concentration of Mn+2 was 10 mM. The binding of Ca+2, Mg+2, Ba+2 or Mn+2 to bovine factor Xa or to the purified coagulant fraction of Russell’s viper venom was very weak in each case.In the absence of Ca+2, the coagulation fraction of Russell’s viper venom could not activate bovine factor X. Activation of factor X was achieved when Ca+2 was replaced by either Mg+2, Ba+2 or Mn+2. When the concentration of these ions were 5 mM, the efficiency of factor Xa generation was estimated to be: Ca+2> Mg+2> Ba+2> Mn+2. Higher concentration of Mg+2, Ba+2, or Mn+2 retarded the activation process. However, Ca+2, Mg+2, Ba+2 or Mn+2 has little or no influence on the esterase activity of factor Xa or purified Rusell’s viper venom.The results suggest that complexation of divalent metal ion with factor X is prerequisite in the activation process. The binding of Mg+2, Ba+2 or Mn+2 to these loosely binding sites might have altered the geometrical configuration as well as the electrostatic environment on factor X significantly. Thus, it is more difficult to form the binary complex and a slower generation of factor Xa results. Therefore, divalent metal ion serves as a dual role in the activation of factor X to factor Xa depending upon the ionic concentration.

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