Zaprinast raises glucose utilisation in SHR skeletal muscle.
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Animals↗
Biomedical subjects
J F Clark
Publications and source records attributed to J F Clark.
ADP-regenerating enzyme systems in mitochondria of guinea pig myometrium and heart.
Any enzyme or enzyme system that produces ADP in proximity to the mitochondria may be capable of stimulating respiration. Hexokinase (HK), adenylate kinase (AK), and mitochondrial creatine kinase (Mi-CK) all catalyze reactions that produce ADP and thus may play a role in cellular nucleotide metabolism or control of mitochondrial oxidative phosphorylation. Respiratory characteristics and enzyme activities of mitochondria simultaneously isolated from heart and uterus of the gravid guinea pig were compared. The abilities of AMP, glucose, and creatine to stimulate mitochondrial respiration via AK, HK, and Mi-CK systems, respectively, were examined. Although the uterine Mi-CK activity is low compared with the values found in heart, the activities of HK and AK were significantly greater. Furthermore, the abilities of HK and AK to stimulate respiration (functional activity) were greater in the uterine mitochondria. Indeed, the activity of AK was sufficient to generate maximal (state 3) respiration. The apparent Michaelis constant (Km) for ADP to stimulate respiration in the isolated uterine mitochondria was significantly different from that of the heart mitochondria (9.6 +/- 0.9 and 5.1 +/- 1 microM ADP, respectively). It is concluded that uterine mitochondria can use HK and AK systems in addition to the CK system in enhancing local ADP concentration, which may aid in the mitochondrial responses to energetic demands.
Adenosine Diphosphate↗
Creatine and phosphocreatine: a review of their use in exercise and sport.
OBJECTIVE: Creatine and phosphocreatine (PCr) are important compounds in the normal energy metabolism of muscle. Recently, it has been shown that dietary creatine (5 to 20 g/day) can increase muscle creatine and PCr, with enhancement in anaerobic exercise performance after two weeks of administration caused by an increase in anaerobic capacity. DATA SOURCES: MEDLINE was searched from 1983 to 1996 using key word "creatine" along with "humans," "muscle," "exercise," and "transport." Also, APStracts, the American Physiology Society search engine for abstracts, was searched from 1994 to 1996. DATA SYNTHESIS: Creatine is transported into the muscle cell by a specific transporter, resulting in increased intracellular creatine and PCr. The PCr is capable of acting as an energy buffer, protecting the adenosine triphosphate (ATP) concentration. Maintaining muscle nucleotides therefore enhances exercise performance and recovery. There have been reports that PCr protects the cells from ischemic damage and decreases the loss of nucleotides by stabilizing cell membranes. Indeed, intravenous PCr (2-4 g/day) has been administered to cyclists, resulting in a faster recovery time between training sessions. CONCLUSIONS/RECOMMENDATIONS: It is becoming evident that oral creatine supplementation may yield certain benefits to enhance the athlete's performance during maximal anaerobic exercise and interval training.
Journal Article↗
A theoretical model of some spatial and temporal aspects of the mitochondrion creatine kinase myofibril system in muscle.
We describe a model of mitochondrial regulation in vivo which takes account of spatial diffusion of high-energy (ATP and phosphocreatine) and low-energy metabolites (ADP and creatine), their interconversion by creatine kinase (which is not assumed to be at equilibrium), and possible functional 'coupling' between the components of creatine kinase associated with the mitochondrial adenine nucleotide translocase and the myofibrillar ATPase. At high creatine kinase activity, the degree of functional coupling at either the mitochondrial or ATPase end has little effect on relationships between oxidative ATP synthesis rate and spatially-averaged metabolite concentrations. However, lowering the creatine kinase activity raises the mean steady state ADP and creatine concentrations, to a degree which depends on the degree of coupling. At high creatine kinase activity, the fraction of flow carried by ATP is small. Lowering the creatine kinase activity raises this fraction, especially when there is little functional coupling. All metabolites show small spatial gradients, more so at low cytosolic creatine kinase activity, and unless there is near-complete coupling, so does net creatine kinase flux. During workjump transitions, spatial-average responses exhibit near-exponential kinetics as expected, while concentration changes start at the ATPase end and propagate towards the mitochondrion, damped in time and space.
Animals↗
The utilisation of creatine and its analogues by cytosolic and mitochondrial creatine kinase.
We have investigated the utilisation of four analogues of creatine by cytosolic Creatine Kinase (CK), using 31P-NMR in the porcine carotid artery, and by mitochondrial CK (Mt-CK), using oxygen consumption studies in isolated heart mitochondria and skinned fibers. Porcine carotid arteries were superfused for 12 h with Krebs-Henseleit buffer at 22 degrees C, containing 11 mM glucose as substrate, and supplemented with either 20 mM beta-guanidinopropionic acid (beta-GPA), methyl-guanidinopropionic acid (m-GPA), guanidinoacetic acid (GA) or cyclocreatine (cCr). All four analogues entered the tissue and became phosphorylated by CK as seen by 31 P-NMR, Inhibition of oxidative metabolism by 1 mM cyanide after accumulation of the phosphorylated analogue resulted in the utilisation of PCr, beta-GPA-P, GA-P and GA-P over a similar time course (approximately 2 h), despite very different kinetic properties of these analogues in vitro. cCr-P was utilised at a significantly slower rate, but was rapidly dephosphorylated in the presence of both 1 mM iodoacetate and cyanide (to inhibit both glycolysis and oxidative metabolism respectively). The technique of creatine stimulated respiration was used to investigate the phosphorylation of the analogues by Mt-CK, Isolated mitochondria were subjected to increasing [ATP], whereas skinned fibres received a similar protocol with increasing [ADP]. There was a significant stimulation of respiration by creatine and cCr in isolated mitochondria (decreased K(m) and increased Vmax vs control), but none by GA, mGPA or beta-GPA (also in skinned fibres), indicating that these latter analogues were not utilised by Mt-CK. These results demonstrate differences in the phosphorylation and dephosphorylation of creatine and its analogues by cytosolic CK and Mt-CK in vivo and in vitro.
Adenosine Diphosphate↗
Increase of flux control of cytochrome c oxidase in copper-deficient mottled brindled mice.
The brindled mottled mouse (Mobr), an animal model of the Menkes' copper deficiency syndrome, was used for the investigation of changes in respiratory flux control associated with cytochrome c oxidase deficiency in muscle. Enzymatic analysis of cardiac and skeletal muscles showed an approximately 2-fold decrease in cytochrome c oxidase activity of brindled mutants in both types of muscles as compared with controls. The activities of NADH-cytochrome c oxidoreductase (respiratory chain segment I-III) and succinate-cytochrome c oxidoreductase (segment II-III) were normal. Assessment of mitochondrial respiratory function was performed using chemically skinned musculus quadriceps or heart muscle fibers isolated from control and brindled mottled mice. In skeletal muscle, there was no difference found in maximal rates of respiration. In the Mobr hearts, this parameter was slightly lower than control. Alternately, the determination of flux control coefficients of cytochrome c oxidase performed by a step by step inhibition of respiration with increasing concentrations of azide or cyanide revealed significantly sharper inhibition curves for brindled mice than for control, indicating more than 2-fold elevated flux control coefficients of cytochrome c oxidase. This investigation proved essential in characterizing the metabolic effect of a cytochrome c oxidase deficiency. We conclude, therefore, that application of metabolic control analysis can be a valuable approach to study defects of mitochondrial oxidative phosphorylation.
Animals↗
Effects of treatment of spontaneously hypertensive rats with the angiotensin-converting enzyme inhibitor trandolapril and the calcium antagonist verapamil on the sensitivity of glucose metabolism to insulin in rat soleus muscle in vitro.
We measured the sensitivity of glucose metabolism to insulin in soleus muscle preparations isolated from spontaneously hypertensive (SH) rats and normotensive age-matched Wistar-Kyoto (WKY) rats. SH rats were treated with the angiotensin-converting enzyme (ACE) inhibitor trandolapril (1 mg/kg) and/or a second antihypertensive drug, the calcium antagonist verapamil, alone (100 mg/kg) or as combination therapy (50 mg/kg). Treatment of SH rats with trandolapril or trandolapril in combination with verapamil for 6 weeks normalized the blood pressure. The estimated concentration of insulin required for half-maximal stimulation of glycogen synthesis (i.e., EC50 values) was approximately 500 microU/ml for muscles from both WKY and SH rats. This value is five times higher than the value obtained from soleus muscle preparations isolated from insulin-sensitive Wistar rats. This indicates that glycogen synthesis is insensitive to insulin in SH and WKY rat soleus muscle. Treatment of SH rats with trandolapril with or without verapamil improved the sensitivity of glycogen synthesis to insulin in soleus muscle. Further experiments investigated whether acute exposure (1 h) of insulin-sensitive skeletal muscle with either trandolaprilat (the active metabolite of trandolapril) or bradykinin (levels of which may be raised by ACE inhibition) could affect the insulin-stimulated rate of glucose metabolism. These results show that both trandolaprilat and bradykinin caused a small but significant increase in the rates of glucose metabolism. In conclusion, 1) SH and WKY rat skeletal muscle was insulin resistant, 2) chronic treatment of SH rats with trandolapril with or without verapamil normalized blood pressure and improved the response of glycogen metabolism to insulin, and 3) bradykinin and trandolaprilat acutely caused a small but significant increase in the rate of glycogen synthesis to a submaximal physiological concentration of insulin.
Angiotensin-Converting Enzyme Inhibitors↗
The creatine kinase equilibrium, free [ADP] and myosin ATPase in vascular smooth muscle cross-bridges.
MgADP has a more pronounced effect on the relaxation behaviour in tonic vascular smooth muscle compared to phasic smooth muscle. An apparent dissociation constant of 1.3 microM has been reported for a high affinity binding site of vascular smooth muscle cross-bridges. For this high affinity to have an effect on the low energy costs of tension maintenance (latch) it would require that free [ADP] in the region of the contractile proteins (at least sometimes) be as low as 1.3 microM. We ask, in this report, whether [ADP] could be as low as 1.3 microM in vascular smooth muscle. If creatine kinase (CK) is at equilibrium, then micromolar [ADP] is incompatible with measured concentrations of phosphocreatine (PCr), free creatine (Cr) and ATP, which entail a mean equilibrium [ADP] of around 18 microM. But CK may not be quite at equilibrium: if there is net PCr synthesis at the mitochondrion, then maintenance of the steady-state requires that there be net PCr hydrolysis in the region of the contractile proteins up to or equal to the rate of myosin ATPase. We derive a simple relationship between net flux and displacement from equilibrium which we use to argue that an [ADP] of 1.3 microM at the contractile proteins would drive significant net PCr synthesis, incompatible with normal contractile function. Thus the CK system holds [ADP] at about 18 microM near the contractile proteins in vascular smooth muscle. We conclude that smooth muscle [ADP] cannot be far from equilibrium and that a role for ADP (at the low micromolar level) in controlling smooth muscle relaxation is unlikely.
Adenosine Diphosphate↗
Metabolite utilization and compartmentation in porcine carotid artery: a study using beta-guanidinopropionic acid.
The relationship between substrate and metabolism in vascular smooth muscle has been investigated by studying the acute energetic effects caused by the creatine analogue beta-guanidinopropionic acid (beta-GPA) on porcine carotid arteries using 31P-nuclear magnetic resonance (NMR). Porcine carotid arteries were superfused for 12 h with Krebs-Henseleit buffer at 22 degrees C, containing 50 mM beta-GPA, and either 11 mM glucose or 5 mM pyruvate as substrate. beta-GPA enters the cells and becomes phosphorylated by creatine kinase to produce beta-GPA-P. Perfusion with beta-GPA leads to the formation of NMR observable beta-GPA-P (after 2.5 h). The appearance of beta-GPA-P with time was significantly greater when glucose was used as substrate. To differentiate between oxidative and glycolytic metabolism in the phosphorylation of beta-GPA, 1 mM cyanide was included in the perfusion buffer containing 50 mM beta-GPA and 11 mM glucose. No phosphocreatine (PCr) was observed with these conditions, and there was a small but significant decrease in ATP concentration ([ATP]) compared with glucose perfusion without cyanide (0.56 +/- 0.02 to 0.47 +/- 0.02 mumol/g wet wt), that was greater than the concentration with pyruvate as substrate (0.25 +/- 0.03 mumol/g wet wt). Thus the [ATP] during cyanide treatment is maintained with glycolytic metabolism. Despite the relatively high [ATP], accumulation of beta-GPA-P only occurred over a much slower time course ( > 10 h) than without cyanide.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenosine Triphosphate↗
Creatine kinase function in mitochondria isolated from gravid and non-gravid guinea-pig uteri.
Mitochondria from gravid and non-gravid guinea pig uteri were isolated and respiratory rates examined to determine the responses to ATP, ADP and creatine. It was found that mitochondria isolated from gravid uterus had (i) a markedly higher respiration rate in state 3; (ii) a greater activation of respiration by creatine in the presence of 0.1 mM ATP and (iii) an elevated specific activity of mitochondrial creatine kinase. It was shown by a competitive enzyme method, using pyruvate kinase to trap ADP, that despite the presence of creatine kinase in the mitochondria, there is no functional coupling between mitochondrial creatine kinase and oxidative phosphorylation as has been shown for striated muscle. It is suggested that the function of uterine Mi-CK is to favour high energy phosphate turnover in conditions of increased metabolic demand in gestating uterine smooth muscle.
Adenosine Diphosphate↗
Actions of the creatine analogue beta-guanidinopropionic acid on rat heart mitochondria.
The action of the creatine analogue beta-guanidinopropionic acid (beta-GPA) was examined in rat heart mitochondria and in isolated cardiomyocytes or fibres which were permeabilized with the non-ionic detergent saponin to determine the kinetics of mitochondrial creatine kinase for beta-GPA. Fibres and myocytes were subjected to increasing [ADP] in the presence and absence of beta-GPA or creatine, whereas isolated mitochondria received a similar protocol with increasing [ATP]. In isolated mitochondria given ATP, there was a stimulation of respiration by creatine, but no significant stimulation of respiration by beta-GPA. Further studies on fibres from control and beta-GPA-fed rats also found that beta-GPA is not utilized by the mitochondria, as evidenced by a lack of beta-GPA-stimulated respiration (Km for ADP = 142 +/- 23 microM) compared with control (Km for ADP from 161 +/- 23 microM), but no significant change in Vmax. Therefore the rat heart mitochondria are not responsive to beta-GPA as compared with creatine. Interestingly, the fibres from beta-GPA-fed rats had no creatine- or beta-GPA-stimulated respiration (Km for ADP = 57.3 +/- 7.2 microM for control, 54.2 +/- 7.2 microM with creatine, and 53.5 +/- 7.8 microM with beta-GPA). The mitochondria prepared from the hearts of rats exposed for 10 weeks to 1% beta-GPA in their diet had a significant decrease in Vmax. and a significant decrease in Km for ADP. Thus the hearts from beta-GPA-fed animals may be pathologic, due to a disruption of the creatine kinase energy circuit.
Adenosine Triphosphate↗
Creatine kinase activity associated with the contractile proteins of the guinea-pig carotid artery.
Activity and role of creatine kinase associated with contractile proteins of vascular smooth muscle have been investigated using skinned guinea-pig carotid artery rings. Membrane solubilization was performed with the detergent Triton X-100. Creatine kinase activity, isoenzyme profile as well as mechanics were performed on the Triton skinned carotid artery rings. Total creatine kinase activity was 47.3 +/- 9.3 IU g-1 ww and electrophoresis showed BB, MB, and MM isoforms (BB-CK being the predominant isoenzyme). One hour incubation with Triton X-100, produced predominantly BB-CK remaining with the myofibrils with some MB, representing 23% of the preskinned creatine kinase activity. When relaxed carotid artery rings were exposed to pCa 9 in the presence of 250 microM ADP, 0 ATP, and 12 mM phosphocreatine, tension was not significantly different from resting tension, but changing to pCa 4.5 caused the carotid artery rings to generate 49.5 +/- 4.5% of maximal tension. When a high-tension rigor state was achieved (250 microM ADP, 0 ATP, 0 phosphocreatine, and pCa 9), the addition of 12 mM phosphocreatine effected significant relaxation. These observations implicate an endogenous form of creatine kinase, associated with the myofilaments, which is capable of producing enough ATP for submaximal tension generation and significant relaxation from rigor conditions. These results suggest co-localization of ATPase, MLCK, and creatine kinase on the contractile proteins of the carotid artery. Such an enzymic association may play a role in the energetic supply to the contractile apparatus of vascular smooth muscle.
Adenosine Diphosphate↗
Alterations in the myocardial creatine kinase system during chronic anaemic hypoxia.
OBJECTIVE: The aim was to use a model of chronic anaemia in the rat, in which there is an increase in cardiac mitochondrial creatine kinase activity (mito-CK) per mitochondrion, to test the hypothesis that creatine stimulated respiration in saponin skinned fibres is correlated with mito-CK activity. In order to discuss the altered regulation of mitochondrial respiratory rate in the context of other metabolic alterations, steady state metabolite concentrations and maximum extracted activities of regulatory enzymes in glycolysis were also investigated. METHODS: Weanling male Wistar Albino rats were randomly distributed into two experimental groups. One group received a powdered diet deficient in iron (5-7 mg iron.kg-1) while the second group was placed on a standard laboratory chow diet (109 mg iron.kg-1) for 4-8 weeks. RESULTS: Total cardiac creatine kinase activity was unchanged in anaemic rats; however, a 25% increase in nascent or functional mito-CK activity per mitochondrion was detected [0.969(SEM 0.005), control group and 1.203(0.040), anaemic group, p < 0.001]. The sensitivity of creatine (40 mM creatine, VCr) and ADP (0.1 mM ADP, V0.1) stimulated respiration, as a percentage of maximum respiratory rate (2.0 mM ADP, V2.0), was increased by 48% and 52% respectively in the anaemic skinned cardiac fibres. An increase in basal respiration with glutamate and malate as substrates was detected in the anaemic group compared to the control group, at 6.77(0.74) v 4.58(0.35) ng O.min-1 x mg-1 dry weight (p < 0.025). Cytosolic ATP was decreased in isolated perfused hearts from anaemic animals, at 35.18(3.11) mumol.g-1 dry weight in control hearts versus 23.66(1.42) in anaemic hearts (p < 0.01). A significant increase in myocardial glycolytic capacity was detected in anaemic cardiac tissue, as evidenced by a 20% increase in phosphofructokinase activity (p < 0.01). Phosphorylase activity was unaltered in anaemic hearts, indicating that the increased glucose requirement originated from exogenous sources. Lactate dehydrogenase (LDH) was increased by 30% in anaemic hearts (p < 0.001). The LDH isozyme profile was shifted in favour of lactate and NAD+ production, thus supporting anaerobic glycolysis. CONCLUSIONS: In support of the phosphocreatine circuit model, the increased mito-CK per mitochondrion in the anaemic skinned fibre preparation was associated with an increase in creatine stimulated respiration. In addition, the sensitivity of mitochondrial respiratory rate to ADP and the maximum glycolytic capacity were increased in anaemic fibres. Although the net effect of these changes in metabolic capacity and regulation on in vivo high energy phosphate flux is unknown, it is likely that they are adaptive alterations that compensate for the lower steady state cytosolic nucleotide concentration.
Adenosine Diphosphate↗
Compartmentation of creatine kinase isoenzymes in myometrium of gravid guinea-pig.
1. This study was performed to investigate the possible presence and role of the creatine kinase (CK) system in the contraction and relaxation of skinned guinea-pig uterus as well as the changes of the CK system during gestation. Experiments were performed on isolated longitudinal fibres of gravid and non-gravid myometrium. 2. Total CK activity increased from 74 +/- 11 to 196 +/- 39 IU (g wet wt)-1 during gestation. 3. The four isoenzymes of CK: muscle (MM), muscle-brain (MB), brain (BB) and mitochondrial (mt-CK) were found in myometrium. MM, MB and BB isoenzymes represented respectively 20.3 +/- 2.6, 10.3 +/- 4.4 and 72.7 +/- 2.2% of total activity. The distribution of isoenzymes did not significantly change with gestation, the contribution of mt-CK increasing from trace to 5% of total activity. 4. BB-CK was specifically bound to Triton X-100-skinned fibres with the non-gravid uterus containing 6.7 +/- 1.9 IU (g wet wt)-1 and the gravid uterus containing 44 +/- 13 IU (g wet wt)-1. 5. Active tension of Triton X-100-treated fibres increased from 6.06 +/- 0.68 to 19.3 +/- 1.9 mM mm-2 during gestation. 6. Submaximal tension (43.3 +/- 4.4% of maximal tension) can be developed in the absence of ATP and in the presence of 12 mM phosphocreatine (PCr) and 250 microM MgADP from endogenous CK in non-gravid uterine fibres while the gravid uterus was able to generate 65.4 +/- 3.9% of maximal tension via the CK system. 7. The endogenous CK system was able to relax the skinned fibres from high-tension rigor conditions by 47.3 +/- 4.2% of total relaxation in non-gravid fibres and 60.6 +/- 3.2% of total relaxation in gravid fibres. 8. Non-gravid and gravid uteri both contained mt-CK of 17.5 +/- 8.4 and 140 +/- 22 micrograms (g wet wt)-1 respectively as determined with antibodies against mt-CK. 9. Oxygen consumption was studied in fibres where the plasmalemma was solubilized with 50 micrograms ml-1 saponin. Maximal respiration was increased from 0.91 +/- 0.05 to 2.61 +/- 0.16 mumol oxygen min-1 (g dry wt)-1 in the gravid uterine fibres. However, creatine did not stimulate respiration in the uterine fibres treated with saponin. 10. It is concluded that the CK system undergoes qualitative as well as quantitative changes during gestation. BB-CK is specifically localized in the myofilaments and mt-CK is present in the uterine mitochondria.(ABSTRACT TRUNCATED AT 400 WORDS)
Actin Cytoskeleton↗
Creatine kinase binding and possible role in chemically skinned guinea-pig taenia coli.
The activity and role of creatine kinase (CK) associated with contractile proteins of smooth muscle have been investigated using skinned guinea-pig taenia coli fibers. Total CK activity was 163 +/- 22 IU/g (ww) and agarose electrophoresis showed BB, MB, and MM isoforms (BB-CK being the predominant isoenzyme). After skinning for 1 h with Triton X-100, BB-CK was specifically associated with the myofibrils, representing 22% of the preskinned CK activity. When relaxed fibers were exposed to pCa 9 in the presence of 250 microM ADP, 0 ATP and 12 mM PCr, tension was not significantly different from resting tension, but changing to pCa 4.5 caused the fibers to generate 59.1 +/- 5.2 percent of maximal tension. When a high-tension rigor state was achieved (250 microM ADP, 0 ATP, 0 PCr, and pCa 9), the addition of 12 mM PCr effected significant relaxation. These observations implicate an endogenous form of BB-CK, associated with the myofilaments and capable of producing enough ATP for submaximal tension generation and significant relaxation from rigor conditions. It was also shown that ADP is bound to the myofibrils and available for rephosphorylation by BB-CK. These results suggest co-localization of ATPase, MLCK and CK on the contractile proteins of the taenia coli. This enzymic association may play a role in the compartmentation of adenine nucleotides in smooth muscle.
Adenosine Diphosphate↗
Amniotic fluid and umbilical artery levels of sex hormones and prostaglandins in human cocaine users.
Biochemical evaluation of amniotic fluid contents is often used to monitor fetal secretory and excretory functions. To determine whether cocaine use during pregnancy affects fetal endocrine secretions, amniotic fluid and umbilical artery samples were collected at term, and selected gonadal hormones and prostaglandins were assayed. Gestational age, birth weights, and placental weights were recorded. The data showed that birth weights were significantly decreased (P less than 0.001) in cocaine users, but gestational age and placental weights were unaffected. Amniotic fluid levels of androstenedione and testosterone were decreased (P less than 0.05) in males born to cocaine users; females were not affected. Prostaglandins (PGF2 alpha and PGE2) were significantly increased (P less than 0.01) in cocaine users. In the umbilical artery, follicle-stimulating hormone was increased (P less than 0.01) in males and females, while luteinizing hormone was increased (P less than 0.01) only in males. We conclude that cocaine passes through the placenta and affects the fetal testes-hypophyseal endocrine system.
Adult↗
The creatine kinase system and cardiomyopathy.
Changes in the creatine kinase system, cellular energetics, regulation of respiration and alterations in parameters of contractility in experimental animals (myopathic hamsters), and in patients with dilated cardiomyopathy were studied. 31P-NMR methods were used to show that cardiomyopathic hearts are characterized by decreased work index, lower tissue ATP, phosphocreatine, and total creatine contents and diminished creatine kinase activity and energy fluxes. In isolated mitochondria, only the creatine kinase activity was decreased. Both in cardiomyopathic hamsters and human hearts a share of mitochondrial creatine kinase in the total tissue enzyme activity was decreased from 33% to 18% and that of BB elevated from 5% in control to 20%, at an unchanged relative level of MM. In saponins-skinned cardiac fibers on cardiomyocytes creatine (Cr, 25 mM) decreased Km for ADP in regulation of respiration from 133 +/- 20 to 20 +/- 4 microM due to activation of coupled mitochondrial creatine kinase-oxidative phosphorylation reactions in control hamster hearts. In the case of cardiomyopathy it decreased Km for ADP only to 81 +/- 13 microM. In endocardial biopsy samples from the hearts of patients with dilated cardiomyopathy taken during angiography, creatine stimulated respiration was decreased by 36% of control value, which correlated well with increase of end-diastolic pressure and fall in ejection fraction. Thus, changes in mitochondrial creatine kinase expression diminished the efficiency of cellular regulation of respiration in cardiomyopathic hearts that may have functional consequences for hemodynamics or may be adaptive alterations in response to decreased contractility.
Animals↗
Multisite saturation transfer using DANTE and continuous wave.
The DANTE pulse sequence was modified to produce selective resonance saturation similar to that produced by the continuous wave (CW) technique. A combined DANTE and CW saturation technique can be used to perform multisite saturation transfer experiments because of the similar saturation produced by the two techniques.
Adenosine Triphosphate↗