scholarly journals Determination of Adenylate Nucleotides in Amphipod Gammarus fossarum by Ion-Pair Reverse Phase Liquid Chromatography: Possibilities of Positive Pressure Micro-Solid Phase Extraction

Separations ◽  
2021 ◽  
Vol 8 (2) ◽  
pp. 20
Author(s):  
Zuzana Redžović ◽  
Marijana Erk ◽  
Ema Svetličić ◽  
Lucija Dončević ◽  
Sanja Gottstein ◽  
...  
Keyword(s):  
Critical Role ◽  
Energy Charge ◽  
Adenosine Monophosphate ◽  
Ion Pair ◽  
Metabolic Energy ◽  
Positive Pressure ◽  
Adenylate Energy Charge ◽  
Reverse Phase ◽  
Gammarus Fossarum

Adenine nucleotides—adenosine monophosphate, diphosphate, and triphosphate—are of utmost importance to all living organisms, where they play a critical role in the energy metabolism and are tied to allosteric regulation in various regulatory enzymes. Adenylate energy charge represents the precise relationship between the concentrations of adenosine monophosphate, diphosphate, and triphosphate and indicates the amount of metabolic energy available to an organism. The experimental conditions of adenylate extraction in freshwater amphipod Gammarus fossarum are reported here for the first time and are crucial for the qualitative and quantitative determination of adenylate nucleotides using efficient and sensitive ion-pair reverse phase LC. It was shown that amphipod calcified exoskeleton impeded the neutralization of homogenate. The highest adenylate yield was obtained by homogenization in perchloric acid and subsequent addition of potassium hydroxide and phosphate buffer to achieve a pH around 11. This method enables separation and accurate detection of adenylates. Our study provides new insight into the complexity of adenylate extraction and quantification that is crucial for the application of adenylate energy charge as a confident physiological measure of environmental stress and as a health index of G. fossarum.

Control of AMP deaminase 1 binding to myosin heavy chain

1998 ◽  
Vol 275 (3) ◽  
pp. C870-C881 ◽  
Author(s):  
Ichiro Hisatome ◽  
Takayuki Morisaki ◽  
Hiroshi Kamma ◽  
Takako Sugama ◽  
Hiroko Morisaki ◽  
...  
Keyword(s):  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Sequence Similarity ◽  
Critical Role ◽  
Energy Charge ◽  
Adenylate Energy Charge ◽  
Amp Deaminase ◽  
Amino Terminal

AMP deaminase (AMPD) plays a central role in preserving the adenylate energy charge in myocytes following exercise and in producing intermediates for the citric acid cycle in muscle. Prior studies have demonstrated that AMPD1 binds to myosin heavy chain (MHC) in vitro; binding to the myofibril varies with the state of muscle contraction in vivo, and binding of AMPD1 to MHC is required for activation of this enzyme in myocytes. The present study has identified three domains in AMPD1 that influence binding of this enzyme to MHC using a cotransfection model that permits assessment of mutations introduced into the AMPD1 peptide. One domain that encompasses residues 178–333 of this 727-amino acid peptide is essential for binding of AMPD1 to MHC. This region of AMPD1 shares sequence similarity with several regions of titin, another MHC binding protein. Two additional domains regulate binding of this peptide to MHC in response to intracellular and extracellular signals. A nucleotide binding site, which is located at residues 660–674, controls binding of AMPD1 to MHC in response to changes in intracellular ATP concentration. Deletion analyses demonstrate that the amino-terminal 65 residues of AMPD1 play a critical role in modulating the sensitivity to ATP-induced inhibition of MHC binding. Alternative splicing of the AMPD1 gene product, which alters the sequence of residues 8–12, produces two AMPD1 isoforms that exhibit different MHC binding properties in the presence of ATP. These findings are discussed in the context of the various roles proposed for AMPD in energy production in the myocyte.

Energy Control During Platelet Secretion:Predominant Role Of ATP-Turnover

1981 ◽  
Author(s):  
A Deijns ◽  
J W N Akkerman
Keyword(s):  
Control Function ◽  
Energy Charge ◽  
Dense Granule ◽  
Metabolic Energy ◽  
Adenylate Energy Charge ◽  
Glucose Addition ◽  
Atp Level ◽  
Atp Turnover ◽  
Dense Granule Secretion ◽  
Granule Secretion

Platelet aggregation and secretion of granular contents require metabolic energy. This implies the existence of a control mechanism that adjusts the rate of energy producing pathways to the energy need of these functions. Such a control function has been attributed to the level of metabolic ATP, to the adenylate energy charge (AEC =(ATP + 1/2ADP)/(ATP + ADP + AMP) and to the velocity of energy generation and consumption (ATP-turnover). In this study we investigated which factor dominates in control of dense granule (3H-serotonin)-, α-granule (β thromboglobulin)- and lysosomal granule (N acetyl β glucosaminidase) secretion. Human gel- filtered platelets were incubated in glucose-free, CN containing medium. Under these conditions ATP-generation only took place in glycogenolysis, which alone was unable to maintain ATP homeostasis. Consequently, metabolic ATP and AEC fell. Addition of glucose restored glycolytic ATP resynthesis wich restored the AEC but the loss of ATP was for the main part irreversible due to hypoxanthin formation. With this system a broad range of metabolic ATP levels and AEC’s could be obtained both at decreasing ATP turnover (before glucose addition) and at increasing ATP turnover (after glucose addition). Analysis of secretion velocity of dense-, α- and lysosomal granules after initiation with thrombin (5 U/ml) showed that at decreasing turnover the secretion velocity of all types of granules depended on both metabolic ATP level and AEC. In contrast, at increasing ATP turnover the correlation between secretion velocity and metabolic ATP level or AEC was lost. Instead, dense granule secretion was faster and lysosomal granule secretion was slower than expected on the basis of metabolic ATP level or AEC, whereas α-granule secretion showed intermediate levels. The data indicate that the three types of granule secretion differ in their dependence on metabolic energy and that apart from metabolic ATP and AEC, ATP turnover is of crucial importance for secretion of granular contents.

Ion-Pair Extraction Cleanup for Liquid Chromatographic Determination of Bentazon in Crops and Soil

1984 ◽  
Vol 67 (3) ◽  
pp. 653-655
Author(s):  
Malin Åkerblom ◽  
Gunborg Alex
Keyword(s):  
Clay Soil ◽  
Chromatographic Determination ◽  
Ion Pair ◽  
Wheat Grain ◽  
Reverse Phase ◽  
Tetrabutylammonium Ion ◽  
Liquid Chromatographic Determination ◽  
Phase Liquid ◽  
Liquid Chromatographic

Abstract Bentazon was selectively extracted as an ion pair with tetrabutylammonium ion into dichloromethane. This technique was used to clean up crop and soil samples before determination of bentazon by reverse phase liquid chromatography and UV detection. Recoveries from potatoes, cucumbers, wheat grain, and clay soil were 77–103%, with a detection limit of 0.02 mg/kg.

Adenylate Energy Charge: A Method for the Determination of Viable Cell Mass in Dental Plaque Samples

1979 ◽  
Vol 58 (4_suppl) ◽  
pp. 2192-2197 ◽  
Author(s):  
Christopher W. Kemp
Keyword(s):  
Computer Program ◽  
Dental Plaque ◽  
Energy Charge ◽  
Cell Mass ◽  
Viable Cell ◽  
Viable Count ◽  
Adenylate Energy Charge ◽  
Interactive Computer Program ◽  
Biochemical Function

The biochemical function, adenylate energy charge (AEC), correlates with the viable count of S. mutans. AEC may be used to estimate the percent viable fraction of bacteria in dental plaque samples. An interactive computer program designed to process the AEC data is described.

Development of an Ion-Pair Reverse-Phase Liquid Chromatographic/Tandem Mass Spectrometry Method for the Determination of an 18-Mer Phosphorothioate Oligonucleotide in Mouse Liver Tissue

2005 ◽  
Vol 11 (2) ◽  
pp. 209-215 ◽  
Author(s):  
Anthony T. Murphy ◽  
Patricia Brown-Augsburger ◽  
Rosie Z. Yu ◽  
Richard S. Geary ◽  
Stefan Thibodeaux ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Liver Tissue ◽  
Mouse Liver ◽  
Ion Pair ◽  
Tandem Mass ◽  
Phosphorothioate Oligonucleotide ◽  
Reverse Phase ◽  
Discovery Research

A quantitative method for the determination of a partially modified, 2′-ribose alkoxy 18-mer phosphorothioate oligonucleotide in liver tissue has been developed. A liquid:liquid extraction, ion-pair reverse phase chromatographic separation and tandem mass spectrometry were used to achieve a quantitation range of 125 to 10,000 ng g−1 mouse liver tissue. A total cycle time of 5 min was obtained while maintaining separation of three potential impurities. Separations were performed using a Discovery RP-Amide C16, 100 × 2 mm column packed with 5 μm particles. The separation was facilitated by the use of triethylamine (TEA) and hexafluoroisopropanol (HFIP) as ion-pair agents. The method has subsequently been used for the determination of other phosphorothioate oligonucleotides in support of discovery research.

Determination of adenine nucleotide levels and adenylate energy charge ratio in two Spartina species

1981 ◽  
Vol 11 ◽  
pp. 37-55 ◽  
Author(s):  
I.A. Mendelssohn ◽  
K.L. McKee
Keyword(s):  
Adenine Nucleotide ◽  
Energy Charge ◽  
Charge Ratio ◽  
Adenylate Energy Charge ◽  
Spartina Species

Reverse-Phase High Performance Liquid Chromatography of Metal Chelates of 4-(2-Pyridylazo)resorcinol and 4-(2-Thiazolylazo)resorcinol. Simultaneous Determination of Low Concentrations of Co, Ni and Fe

1994 ◽  
Vol 59 (10) ◽  
pp. 2209-2226 ◽  
Author(s):  
Josef Doležal ◽  
Lumír Sommer
Keyword(s):  
High Performance Liquid Chromatography ◽  
High Performance ◽  
Cetyltrimethylammonium Bromide ◽  
Ion Pair ◽  
Metal Chelates ◽  
Reverse Phase ◽  
Rp Hplc ◽  
Low Concentrations ◽  
Methanol Solutions

The behaviour of stable and inert metal chelates of 4-(2-pyridylazo)resorcinol and 4-(2-thiazolylazo)resorcinol in RP HPLC and in its ion-pair modification (IP RP HPLC) on Separon SGX RPS silica gel was studied. Good results were obtained by the ion-pair variant in water methanol solutions at pH 7 in the presence of cetyltrimethylammonium bromide. The technique proved to be convenient for the preconcentration, separation and quantitation of low concentrations of Fe, Co, and Ni in waters.

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