• Title/Summary/Keyword: creatine

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Overexpression, Purification, and Preliminary X-ray Crystallographic Analysis of Human Brain-Type Creatine Kinase

  • Bong, Seung-Min;Moon, Jin-Ho;Jang, Eun-Hyuk;Lee, Ki-Seog;Chi, Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.295-298
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    • 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%.

Evidences for Functionally Direct Coupling between Mitochondrial Phosphocreatine Formation and Oxidative Phosphorylation (Oxidative Phosphorylation계에 의한 Phosphocreatine의 생성 기작)

  • Kim, Il-Han
    • The Journal of Natural Sciences
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    • v.5 no.2
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    • pp.3-11
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    • 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.

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Effects of short term creatine loading on performance, plasma components and anaerobic power in taekwondo player (단기간의 크레아틴 섭취가 태권도 선수의 운동수행 능력, 혈장요소 및 무산소성 파워에 미치는 영향)

  • Kim Sun-Ho;Ku Min;Min Bome-Il;Lee Hong-Min;Ko Young-Ho;Yoon Young-Bok
    • Proceedings of the Korea Contents Association Conference
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    • 2005.05a
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    • pp.191-200
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    • 2005
  • The main purpose of the present study was to find the effect of short term creating loading on repeated bouts of kicking, plasma creating phosphokinase and anaerobic power in male taekwondo player from high school. Sixteen male were randomly assigned to two groups. They were tested before and after 6 days of placebo$(4{\times}5g\;glucose\;d^{-1},\;N=11)$ or Creatine monohydrate loading$(consume\;0.3g\;kg^{-1},\;N=11)$. Repeated bouts of kicking was tested through maximal front kicking continuously for 1 minute and took a rest for 30 seconds, after 30 seconds rest they should have to start kicking again. They repeated it 2 times more(total 3 sets) and the kicking times were measured each sets. Also, body composition, CPK, mean power and peak power were measured before and after experiment. The following results were obtained from this study; 1. The kicking test was significantly increased at 1, 2, 3 sets only in creatine loading group(p<.05, p<.01). 2. Only creatine loading group was significantly increased of plasma creatine phosphokinase(p<.01). 3. Only creatine loading group were significantly increased both of mean power and peak power(p<.05; p<.01). These results suggest that short creatine loading was effective diet protocol in taekwondo player from male high school.

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Simultaneous Determination of Creatine, Dicyandiamide and Dihydrotriazine in Dietary Supplements by High Performance Liquid Chromatography (고성능 액체크로마토그래피를 이용한 식이보충제에서 크레아틴, 디시안디아마이드, 디하이드로트리아진의 동시분석)

  • Park, Sang-Wook;Yoo, Myung-Sang;Lee, Wonjae
    • KSBB Journal
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    • v.29 no.4
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    • pp.232-238
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    • 2014
  • The simultaneous determination of creatine monohydrate (CrM), dicyandiamide and dihydrotriazine in dietary supplements using reversed-phase high performance liquid chromatography (HPLC) was developed. Chromatography was performed on a Nuclosil 100-5 SA ($4.6{\times}250mm$) column with a mobile phase of 2.3% ammonium phosphate (pH 5.5), and UV detection at 224 nm, 212 nm, and 237 nm, respectively. The performance characteristics of HPLC were determined in terms of selectivity, linearity, precision, recovery, limit of detection (LOD), and limit of quantification (LOQ). The calibration curves were linear within the concentration range of $40.0{\sim}500.0{\mu}g/mL$ for creatine, $0.1{\sim}12.8{\mu}g/mL$ for dicyandiamide, and $0.05{\sim}6.4{\mu}g/mL$ for dihydrotriazine. The detection limits of the method were 1.09, 0.01, and $0.08{\mu}g/mL$ for creatine, dicyandiamide, and dihydrotriazine, respectively. The recoveries of creatine, dicyandiamide, and dihydrotriazine were 97.2~100.9, 92.3~106.5, and 97.2~105.5%, respectively. It is expected that the chromatographic analytical method developed in this study will be usefully applicable to simultaneous determination of creatine, dicyandiamide, and dihydrotriazine contained in dietary supplements.

Immediate Decrease in γ-AminoButyric Acid after Caffeine Intake in Adolescents: a Preliminary MRS Study

  • Hahn, Sanghyun;Kim, Yun Ho;Seo, Hyung Suk
    • Investigative Magnetic Resonance Imaging
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    • v.21 no.2
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    • pp.102-105
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    • 2017
  • In adolescents, sleep deprivation problem is getting worse, and increased caffeine consumption is considered to relieve the stress caused by sleep deprivation and academic burden. In this study, immediate neurologic effects of caffeine intake on adolescents were evaluated in three high school students using the ${\gamma}-aminobutyric$ acid (GABA)/creatine ratio on magnetic resonance spectroscopy (MRS). MEGA-PRESS MRS and TE 135 ms single voxel MRS were performed in the anterior cingulate cortex before and after drinking a cup of coffee, which contained 104 mg of caffeine. GABA and creatine were measured on LCModel 6.3, respectively. In all three students, GABA/creatine ratios were decreased after caffeine intake. The GABA/creatine ratios obtained before caffeine intake were decreased after caffeine intake in all the three adolescents. In this preliminary study, caffeine intake caused an immediate decrease in the GABA/creatine ratio in the brain and it may be related to the neurologic effects of caffeine on an adolescent's brain.

The Effect of $Mg^#$, $Ca^#$, $Na^+$, $K^+$ and Creatine Phosphate on the ATPase Activity of Microsomal Fraction from Rabbit Uterus (가토자궁근(家兎子宮筋)에서 분리(分離)한 Microsome 분획내(分劃內) Adenosinetriphosphatase 활성도(活性度)에 미치는 $Mg^#$, $Ca^#$, $Na^+$, $K^+$ 및 Creatine phosphate의 영향(影響))

  • Choi, Sin-Jyoung
    • The Korean Journal of Pharmacology
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    • v.2 no.2
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    • pp.35-42
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    • 1966
  • The author investigated the effect of $Mg^#$, $Ca^#$, $Na^+$, $K^+$ and creatine phosphate on the ATPase activity of microsomal fraction isolated from rabbit uterus and obtained the following results : 1) The uterine microsomal fraction contained the $Na^+-$ and $K^+-$ activated ATPase in the presence of $Mg^#$. The ATPase activity increased with protein content in the fraction. 2) The maximum ATPase activity was obtained at $Na^+$ and $K^+$ concentraction of 100 mM respectively. 3) In the absence of $Mg^#$, the ATPase was not activated by $Na^+$ and $K^+$, but inhibited. 4) Car stimulated the $Na^+-$ and $K^+-$ activated ATPase in the presence of $Mg^#$. However, in the absence of $Mg^#$, the ATPase was not activated by $Ca^#$. 5) The $K^+-$ activated ATPase activity was greater than the $Na^+-activated$ ATPase under all conditions. 6) The $Na^+-$ and $K^+$ activated ATPase activity was increased by addition of creatine phosphokinase and creatine phosphate to the reaction mixture.

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The Effects of Repetitive Exercise on the Blood Cortisol, MDA, and Creatine Kinase Activity in Judoist. (유도선수들의 반복운동이 혈중 코티졸과 지질과산화 및 creatine kinase 활성에 미치는 영향)

  • 백일영;곽이섭;이문열
    • Journal of Life Science
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    • v.14 no.4
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    • pp.573-576
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    • 2004
  • The purpose of the present study is to investigate the lipid peroxidation, creatine kinase activity and cortisol hormone levels following the training intensity in elite judo players. Six elite Judo players participated in the experiments (3h repetition judo program), which include stretching, judo skill practice and cool down without recess. Blood sampling were taken at the judo gymnasium at the time of resting, 1h training, 2h training, 3h training, 2h recovery, and 24h recovery time and this were analyzed for CK, MDA and Cortisol levels. The results obtained were analyzed via repeated measures of ANOVA using SPSS package program (ver.10.0) and a value of p<.05 was considered statistically significant. The results from this study were as follows. In the CK levels, which reflect the contribution of creatine phosphate and muscle damage degree, there was a significant difference (p<.05) after judo training in every period. Recovery 24h showed the highest level. In the MDA levels, which reflect lipid peroxidation, there was a significant difference (p<.05) after judo training. Recovery 2h showed the lowest level. In the cortisol hormone levels, which reflect stress status, there was a significant difference (p<.05). In this study, we can conclude that For the trained athletes, MDA level was lower at the time of exercise compare to the other period, this is caused by the increased antioxidant defence mechanism.

Guanidinoacetate Methyltransferase Expression in Mouse Epididymis (마우스 부정소에서 Guanidinoacetate Methyltransferase의 발현)

  • Lee, Hang;Lee, Mun-Han
    • Development and Reproduction
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    • v.1 no.2
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    • pp.117-124
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    • 1997
  • Guanidinoacetate N-methyltransferase (GAMT) catalyzes the last step of creatine biosynthesis and the resultant creatine plays an important role in cellular energy metabolism. GAMT is mainly found in liver, kidney as well as testis and epididymis. We have localized the site of creatine biosynthesis in mouse epididymis by immunoperoxidase staining of GAMT using anti-GAMT antibody. Gamt is extensively expressed in the microvilli of epididymal epithelial cells and also expressed weakly in the cyto plasm of the cells. The staining of GAMT was most prominent in the microvilli of proximal caput epididymis and the intensity was progressively diminished as the epididymal tubule proceeds toward caudal part. The result suggests that GAMT or Cr might be involved in sperm function and/or maturation process in epididymis.

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Enhanced Myocardial Protection by Addition of Creatine Phosphate to the St. Thomas Hospital Cardioplegic Solution -Studies in the rat - (St. Thomas Hospital 심정지액에 Creatine Phosphate 를 첨가한 후 심근 보호 효과)

  • 최순호
    • Journal of Chest Surgery
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    • v.22 no.4
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    • pp.580-588
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    • 1989
  • The potential for enhancing myocardial protection by adding high-energy phosphate to cardioplegic solutions [St. Thomas Hospital solution] was investigated in a rat heart model of cardiopulmonary bypass and ischemic arrest. Creatine phosphate was evaluated as an additive to the St. Thomas Hospital cardioplegic solution. Creatine phosphate 10.0 mmol/L as the optimal concentration which improved recovery of aortic flow and cardiac output after a 30 minute period of normothermic [37oC] ischemic arrest. In comparing mechanical function in both groups the mean postischemic recoveries of aortic flow, cardiac output, stroke volume and stroke work [expressed as a percentage of its preischemic control] were significantly greater in STH-CP group than in CP- free control group. In addition to improving function and decreasing CK release, CP reduced reperfusion arrhythmias significantly decreasing the time between cross-clamp removal and return to regular rhythm from 81.8 * 13.9 [sec] in CP-free group to 35.9 * 6.8 [sec] in CP group [P< 0.05] so, exogenous CP exerts potent protective and antiarrhythmic effects when added to the St. Thomas Hospital cardioplegic solution. However, the mechanism of action remains to be elucidated.

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Creatine Kinase and its Isoenzymes in Jindo Dogs (진도견의 creatine kinase 활성치와 isoenzyme)

  • 최석화
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.41-45
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    • 1998
  • This study was performed to investigate the serum creative kinase(CK) activity and CK isoenzyme in Jindo dog. Serum CK activity and CK-isoenzyme were analyzed in 53 Jindo dogs of both sexes. The mean value and normal range of serum CK activity were 24.1 lu/$\ell$ and 7-91 lu/$\ell$, respectively, in 29 female dogs, 24.8 lu/$\ell$ and 8-89 lu/$\ell$ in 24 male dogs. The CK activity of the Puppy showed a tendency to be higher than that of the adult. There was no significance between Puppy and adult. Three isoenzymes (CK-MM, CK- BB, and CK-MB) were recognized in serum. The mean percentages of female and male were as follows: 48.31fp and 48.1% for CK-MM, 35.49) and 33.61fp for CK-BB, and 8.2% and 10.1% for CK-MB in the puppy and 46.21% and 46.1 % for CK-MM, 36.3% and 37.6% for CK-BB and 10.5% and 9.5% for CK-MB in the adult.

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