• The Journal of Natural Sciences
• /
• v.5 no.2
• /
• pp.3-11
• /
• 1992

As concentration of ADP was increased, the rate of phosphocreatine formation by respiring heart mitochondria was increased. The value of apparent Km of the phosphocreatine-forming mitochondria for ADP was estimated to be 0.0185 mM. This value was much lower than that of Km for ATP (0.31 mM) which was determined from the reaction of the soluble form of mitochondrial creatine kinase. The concentration of ATP remained constant during the respiring in the presence of ADP. The rate of accumulation of oxidative-phosphorylated ATP in the mitochondrial respiration medium was continuously monitored as a function of ADP concentration with the firefly luciferase-coupled assay. In that case, exogenous creatine did not affect the rate of accumulation of ATP, indicating that phosphocreatine-forming (i.e.,respiring) mitochondria in the presence of ADP did not use the ATP in the medium as a substrate.These results suggest that the heart mitochondrial creatine kinase bound to the inner membrane functionally tight-coupled to the oxidative phosphorylating system with respect to the respired ATP.

• Korean Journal of Veterinary Research
• /
• v.35 no.2
• /
• pp.263-270
• /
• 1995

Creatine(Cr) and phosphocreatine(PCr), the important mediators of intracellular high-energy phosphate buffer system, were found in the tissues of mouse seminal vesicle and also in the extracellular fluids of seminal vesicle secretion. This study was performed m confirm that the secretion and accumulation of Cr and PCr is regulated by testosterone and its $5{\alpha}$-reduced metabolite, $5{\alpha}$-dihydrotestosterone(DHT). In addition, the effect of nandrolone decanoate(ND), a synthetic anabolic steroid, on the levels of Cr and PCr in the seminal vesicle was compared with those of testosterone propionate(TP) and DHT. Male Swiss-Webster mice were castrated and three groups of the castrates were treated with daily injection(sc) of same molar dose($1.45{\times}10^{-8}mol/g\;BW$) of TP, DHT, or ND. All three androgens rapidly increased weights of seminal vesicle tissue and fluid, and also increased concentrations of Cr and PCr in the tissue and fluid. However, ND was least effective in increasing seminal vesicle weights, whereas ND was as effective as, or in some cases, more effective than, TP or DHT in increasing Cr and PCr levels in the tissue and fluid. The results confirm that the accumulation of Cr and PCr in the seminal vesicles is regulated by testosterone and DHT, and also suggest that the effects of androgens on seminal vesicle growth and secretory activity may be differentiated.

• Proceedings of the KSMRM Conference
• /
• 2003.10a
• /
• pp.84-84
• /
• 2003

Purpose： To assess clinical proton MR spectroscopy (MRS) as a noninvasive method for evaluating tumor malignancy at 3T high field system. Materials and methods： Using 3T MRI/MRS system, localized water-suppressed single-voxel technique in patients with brain tumors was employed to evaluate spectra with peaks of N-acetyl aspartate (NAA), choline-containing compounds (Cho), creatine/phosphocreatine (Cr) and lactate. On the basis of Cr, these peak areas were quantificated as a relative ratio.

• Korean Journal of Exercise Nutrition
• /
• v.16 no.1
• /
• pp.35-41
• /
• 2012

We investigated the role of fatty acid availability on skeletal muscle AMPK activity and adenine nucleotide content. To investigate the chronic effects of elevated fatty acid in vivo Sprague-Dawley rats were fed a chow diet (15% fat) or a diet high in saturated (SAFA, 52% fat) or polyunsaturated (PUFA, 52% fat) fat for eight weeks. High fat diets increased (P < 0.05) plasma FFA levels by 25%. AMPK activity was increased in SAFA and PUFA rats and occurred in the absence of changes in ATP, AMP, phosphocreatine and glycogen content. These results suggest that increasing fatty acid availability increases AMPK activity independent of changes in the cellular energy charge, and implicate the regulation of AMPK by a covalent mechanism. These data also support the contention that increasing fatty acid availability can increase subsequent fatty acid oxidation by an AMPK-mediated process.

• Journal of Ginseng Research
• /
• v.44 no.3
• /
• pp.461-474
• /
• 2020

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, the mechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides) in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potential target in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides, had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol (PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in the study, was selected as a representative to confirm direct binding and its biological importance. Biolayer interferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPD specifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by molecular docking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activity in vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the function of the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delaying exercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginseng reduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can help in further developing better CK-MM activators based on the dammarane-type triterpenoid structure.

• Proceedings of the Korean Society of Medical Physics Conference
• /
• 2003.09a
• /
• pp.76-76
• /
• 2003

Purpose: To assess clinical proton MR spectroscopy (MRS) as a noninvasive method for evaluating tumor malignancy at 3T high field system. Methods: Using 3T MRI/MRS system, localized water-suppressed single-voxel technique in patients with brain tumors was employed to evaluate spectra with peaks of N-acetyl aspartate (NAA), choline-containing compounds (Cho), creatine/phosphocreatine (Cr) and lactate. On the basis of Cr, these peak areas were quantificated as a relative ratio. Results: The variation of metabolites measurements of the designated region in 10 normal volunteers was less than 10%. Normal ranges of NAA/Cr and Cho/Cr ratios were 1.67$\pm$018 and 1.16:1:0.15, respectively. NAA/Cr ratio of all tumor tissues was significantly lower than that of the normal tissues (P=0.005). Cho/Cr ratio of high-grade gliomas was significantly higher than that of low-grade gliomas (P= 0.001), Except 4 menigiomas, lactate signal was observed in all tumor cases. Conclusions: The present study demonstrated that the neuronal degradation or loss was observed in all tumor tissues. Higher grade of brain tumors was correlated with higher Cho/Cr ratio, indicating a significant dependence of Cho levels on malignancy of gliomas. This results suggest that clinical proton MR spectroscopy could be useful to predict tumor malignancy. Acknowledgement: This study was supported by a grant of the Mid and Long Term Nuclear R/D Plan Program, Ministry of Science and Technology, Republic of Korea.

• Proceedings of the Korean Society of Medical Physics Conference
• /
• 2003.09a
• /
• pp.73-73
• /
• 2003

Purpose: To determine the motor cortex dysfunction in hemiparetic patients due to deep intracerebral hematoma, authors performed proton magnetic resonance spectroscopy (lH MRS) for the evaluation of biochemical changes in the cortex on affected hemisphere according to axonal injury at the level of internal capsule. Methods: Ten control subjects and 14 patients with documentable hemiparesis of varying severity hemiparesis were included. All the hemiparesis was caused by deep intracerebral hematoma (putaminal and thalamic hemorrhage). In vivo 1H MRS study was performed on a 3T MRI/MRS system using STEAM sequence. As a single-voxel technique, Spectral parameters were: 20 ms TE, 2000 ms TR, 128 averages, 2500 Hz spectral width, and 2048 data points. Results: We found that the mean N-acetylaspartate (NAA)/phosphocreatine (Cr) and NAA/choline (Cho) ratios were significantly decreased in the motor cortex of the hemiparesis patients compared with control subjects. Conclusions: 1H MRS examinations of the motor cortex might help to differentiate distinct clinical entities of hemiparesis and to monitor pharmacological effects in therapeutic trials, providing a quantitative biological marker for motor neuron dysfunction. Acknowledgement: This study was supported by a grant of the Center for Functional and Metabolic Imaging Technology, Ministry of Health & Welfare, Republic of Korea. (02-PJ3-PG6-EV07-000).

• Journal of the Korean Magnetic Resonance Society
• /
• v.15 no.1
• /
• pp.54-68
• /
• 2011

$^1H$ nuclear magnetic resonance (NMR) spectroscopy of biological samples has been proven to be an effective and nondestructive approach to probe drug toxicity within an organism. In this study, ketoprofen toxicity was investigated using $^1H$-NMR spectroscopy coupled with multivariate statistical analysis. Histopathologic test of ketoprofen-induced acute gastrointestinal damage in rats demonstrated a significant dose-dependent effect. Furthermore, principal component analysis (PCA) derived from $^1H$-NMR spectra of urinary samples showed clear separation between the vehicle-treated control and ketoprofen-treated groups. Moreover, PCA derived from endogenous metabolite concentrations through targeted profiling revealed a dose-dependent metabolic shift between the vehicle-treated control, low-dose ketoprofen-treated (10 mg/kg body weight), and high-dose ketoprofen-treated (50 mg/kg) groups coinciding with their gastric damage scores after ketoprofen administration. The resultant metabolic profiles demonstrated that the ketoprofen-induced gastric damage exhibited energy metabolism perturbations that increased urinary levels of citrate, cis-aconitate, succinate, and phosphocreatine. In addition, ketoprofen administration induced an enhancement of xenobiotic activity in fatty oxidation, which caused increase levels of N-isovalerylglycine, adipate, phenylacetylglycine, dimethylamine, betaine, hippurate, 3-indoxylsulfate, N,N-dimethylglycine, trimethyl-N-oxide, and glycine. These findings demonstrate that $^1H$-NMR-based urinary metabolic profiling can be used for noninvasive and rapid way to diagnose adverse drug effects and is suitable for explaining the possible biological pathways perturbed by nonsteroidal anti-inflammatory drug toxicity.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.2
• /
• pp.295-298
• /
• 2008

Creatine kinase (CK; E.C. 2.7.3.2) is an important enzyme that catalyzes the reversible transfer of a phosphoryl group from ATP to creatine in energy homeostasis. The brain-type cytosolic isoform of creatine kinase (BB-CK), which is found mainly in the brain and retina, is a key enzyme in brain energy metabolism, because high-energy phosphates are transfered through the creatine kinase/phosphocreatine shuttle system. The recombinant human BB-CK protein was overexpressed as a soluble form in Escherichia coli and crystallized at $22^{\circ}C$ using PEG 4000 as a precipitant. Native X-ray diffraction data were collected to $2.2{\AA}$ resolution using synchrotron radiation. The crystals belonged to the tetragonal space group $P4_32_12$, with cell parameters of a=b=97.963, $c=164.312{\AA},\;and\;{\alpha}={\beta}={\gamma}=90^{\circ}$. The asymmetric unit contained two molecules of CK, giving a crystal volume per protein mass $(V_m)$ of $1.80{\AA}^3\;Da^{-1}$ and a solvent content of 31.6%.

• Journal of the Korean Electrochemical Society
• /
• v.15 no.4
• /
• pp.249-255
• /
• 2012

The planar-type amperometric creatinine biosensor employing an additional enzyme layer containing creatine kinase and adenosine triphosphate was developed to eliminate severe interference from creatine. In the additional enzyme layer, an interfering substance, creatine is converted to noninterfering product, phosphocreatine. Furthermore, the carbon electrode electroplated with Pt black(Pt-B) was employed to fabricate creatinine biosensors with improved sensor performance(e.g., sensitivity, reliability, and reproducibility). The creatinine levels in an unknown sample were determined within less than 5% errors using creatinine microsensors equipped in a flow-cell cartridge.