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The biosensor can be used to measure micromolar concentrations of GDP on the background of millimolar concentrations of GTP.
The concentrations of GDP were measured by the absorbance at 260 nm using an extinction coefficient of 13,804 M−1 cm−1 [ 18].
Concentrations of GDP and GMP showed a similar behavior as those of ADP and AMP, i.e. concentrations increased (data not shown).
We have also shown for the first time that the presence of EF-Ts shifts the K M for the interaction of EF-Tu with its substrate GDP allowing EF-Tu to function at significantly lower concentrations of GDP.
In practice, the biosensor is able to detect down to 2 μM concentrations of GDP in the presence of mM GTP, as demonstrated in the GTPase and nucleotide exchange assays.
The magnitude of negative cooperativity between agonists and GDP in these experiments was derived from a decrease in displacement of [H]NMS binding by a fixed concentration of tested agonist by increasing concentrations of GDP in GDP-less membranes.
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All in vitro studies have been done with extremely high concentrations of GDPs, compared with their concentrations in peritoneal dialysis solutions.
Time courses show that GDP binding is fast, completed in <1 s even at the lowest concentration of GDP.
In the chemical equilibrium between the RC RF1RF3 complex and the RC RF1 complex accompanied by free RF3GDP, the former complex dominates at low concentrations of free GDP, whereas the latter two dominate at high concentrations of free GDP.
The mean and standard deviation of guanine and pyridine nucleotides concentrations are presented in Table 2 There were no significant changes in the concentrations of GTP, GDP, GMP and TGN in red blood cells immediately after exercise, and at the 30th min of recovery.
We then incubated the oligomers with high concentrations of Cdc42-GDP and looked for additional densities corresponding to Cdc42-GDP by EM and SPA.
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