Rapid Adenosine Research Articles

ATP Screening Method for Presumptive Detection of Microbiologically Contaminated Carbonated Beverages

ABSTRACTA rapid adenosine triphosphate (ATP) screening method utilizing a vacuum‐filtration procedure was developed for the presumptive detection of yeast and mold contamination in carbonated beverages. First, a baseline of ATP content was established by examining 240 retail cola samples that did not contain detectable microorganisms. Then an additional 182 retail samples were assayed for ATP and Colony Forming Units (CFU). Of these, less than 3% were presumptive‐positive by the ATP method. All 182 samples were microbiologically negative for yeast. When 140 cola samples were spiked with various yeast types, more than 89% of samples were correctly identified as presumptive‐positive at an arbitrary product specification level of 5 CFU/mL.

Activation of the dynein adenosinetriphosphatase by microtubules.

Previous work has indicated that following the rapid adenosine 5'-triphosphate (ATP) induced dissociation of the microtubule-dynein complex, the rate-limiting step in the ATPase cycle is product release [Johnson, K. A. (1983) J. Biol. Chem. 258, 13825-13832], which occurs at a rate of approximately 2-6 s-1. In this report we complete the analysis of the ATPase cycle by examining the effect of microtubules on the rate of product release. For these studies we used repolymerized Tetrahymena axonemal microtubules and microtubule-associated protein (MAP) free bovine brain microtubules which were shown to be free of any measureable ATPase activity. Tetrahymena 22S dynein bound to these microtubules predominantly by the ATP-sensitive site and at a rate giving an apparent second-order rate constant of (0.2-1) X 10(6) M-1 s-1, which is 50-fold greater than the rate observed with brain microtubules containing MAPs. ATP induced the rapid dissociation of the microtubule-dynein complex with an apparent second-order rate constant vs. ATP concentration equal to 1.6 X 10(6) M-1 s-1; this value is only slightly lower than that observed in the presence of MAPs. After the ATP-induced dissociation, the dynein reassociated with the microtubules following a lag period due to the time required to hydrolyze the ATP. The duration of the lag time for reassociation decreased with increasing microtubule concentration, suggesting that microtubules increased the rate of ATP turnover. Direct measurements at steady state showed that the specific activity of the dynein increased with increasing microtubule concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Some biochemical properties of the rapid adenosine uptake system in rat brain synaptosomes.

The rapid uptake of adenosine into rat brain cortical synaptosomes is mediated by a facilitated diffusion process. The carrier mediated uptake is sensitive to pH and temperature. The average Q10 value for the system is approximately 1.77 and the necessary activation energy (Ea) is estimated to be 8870 cal/mol. These values are essentially in agreement with values reported for adenosine uptake carriers of other tissues. Substrate specificity of the uptake system in the CNS demonstrates that nucleotides do not interact with the carrier until they have been hydrolyzed to nucleosides. Structural modification of the purine moiety at the "2" position did not have a profound effect on the ability of the molecule to serve as a substrate for the uptake system. Competitive inhibition by sulfhydryl reagents, p-chloromercuribenzoate, and N-ethylmaleimide on adenosine uptake suggests a direct involvement of sulfhydryl group(s) in the uptake mechanism. Other purines and pyrimidines also inhibit adenosine uptake, suggesting that a variety of nucleosides can interact with a common carrier system.

Uptake and metabolism of adenosine by human erythrocyte ghosts.

Uptake and metabolism of adenosine by human erythrocyte ghosts.